Novel cephalosporin derivatives and pharmaceutical compositions thereof

ABSTRACT

The present invention relates to novel cephalosporin derivatives represented by Chemical Formula 1. 
     
       
         
         
             
             
         
       
     
     Wherein, X, Y, L, R 1 , and R 2  are as same as defined in the description of the invention. 
     The present invention also relates to pharmaceutical antibiotic compositions comprising a novel celphalosporin derivative represented by Chemical Formula 1, a prodrug thereof, a hydrate thereof, a solvate thereof, an isomer thereof, or a pharmaceutically acceptable salt thereof as an effective ingredient. 
     According to the present invention, novel cephalosporin derivatives, a prodrug thereof, a hydrate thereof, a solvate thereof, an isomer thereof, or a pharmaceutically acceptable salt thereof as an effective ingredient for the broad spectrum of antibiotic resistant, low toxicity, particularly in Gram-negative bacteria, which can be useful with strong antimicrobial activity.

CROSS-REFERENCE TO RELATED APPLICATION

This application is a continuation of PCT International ApplicationSerial No. PCT/KR2012/002302 filed Mar. 29, 2012, which claims thebenefit of Korean Patent Application No. 10-2011-0028603, filed Mar. 30,2011, the contents of each of which are incorporated herein byreference.

TECHNICAL FIELD

The present invention relates to novel cephalosporin derivatives. Thepresent invention also relates to pharmaceutical antibiotic compositionsincluding novel cephalosporin derivatives, prodrug thereof, hydratethereof, solvates thereof, isomers thereof or pharmaceuticallyacceptable salts thereof as an effective ingredient.

BACKGROUND

The treatment of Gram-negative bacteria has intensified with manydevelopment programs during its golden era from the 1960s through to the1980s. However, with the increasing Gram-positive bacteria infections,such as MRSA (Methicillin-resistant Staphylococcus aureus) in the 1990s,the Gram-negative researches were overshadowed. Since late 2000, due tothe growing concern of the lack of multidrug-resistant Gram-negativebacteria treatment, the Gram-negative bacteria research re-gained itsinterest. According to the recent publication by the Infectious DiseasesSociety of America (IDSA), The European Centre for Disease Preventionand Control (ECDC), and the European Medicines Agency (EMEA), there areonly 8 effective drugs against Gram-negative bacteria worldwide.Especially the multidrug-resistant Gram-negative bacteria new drugdiscovery is extremely scarce.

In particular, the recently discovered NDM-1 (New Delhimetalo-beta-lactamase) is rapidly spreading and became a threat tointernational community. The NDM-1 mainly appears in Gram-negativebacteria, and currently colistin and tigecycline are the only twoeffective drugs. However, these drugs are not readily used due to theirtoxicity and side effects. Thus urgent needs to replace these two drugsare in demand. The rapid spread of these pathogens is not just a burdenon few affected countries but on every country where an internationaljoin effort is a must to control such spread.

Already in 2004, the Infectious Diseases Society of America (IDSA) hadpublished a report called the “Bad Bugs, No Drugs.” A hit list waspublished in this report as the current global rate of resistanceincreases. The list is based on the morbidity, mortality with highpathogen, and absence of the effective drug therapy. Amongst the list, 3of them are the Gram-negative bacteria: P. aeruginosa, A. baumannii, andK. pneumoniae isolates. They require government's support as they createserious disease outbreak problems. Currently, there are few classes ofdrug available against these bacteria, such as cephalosporins,carbapenems, aminoglycosides, and tigecyclines. However, there are noeffective drugs available against the resistant stains, and especiallyagainst acinetobacter, tigecyclines is the only effective class of drug.

In 2006, the multidrug-resistant K. pneumoniae was reported in patientswith XDR-KP only in the eastern part of the United States, but morerecently, it spread throughout the rest of the country. In case ofacinetobacter, the infection spread nationwide by the soldiers who werepreviously deployed to Middle Eastern countries. Carbapenems is mainlyused as the leading treatment, but there is rapid increase incarbapenems resistant stains, thus it is left with any effectivetreatment.

As the demand increases for Gram-negative bacteria treatments,pharmaceutical companies are showing strong interest, but only fewantibiotics are in development. Among them is β-lactam inhibitors, andsome noteworthy compounds are CEF-104 and CAZ-104 from Novexel, CAX-201from Cubist, and one compound from each of the following classes:Polymyxin, tetracycline and aminoglycoside. Among effectiveacinetobacters, there are PTK-0796, a tetracycline class, andCB-182,804, a polymyxin derivative. However, these two compounds are notwidely used due to their toxicity issues in their safety profile.

Currently cephalosporin and carbapenem are the two most widely usedGram-negative antibiotics classes. Within carbapenem class, imipenem andmeropenem are the market dominating compounds, but the predominantmarket leading compounds are the generic drugs. Ceftobiprole was themost promising candidate within the cephalosporin class, butunfortunately its development program was discontinued. Therefore,within the cephalosporin class, generic compounds and combi-therapy willbe the main treatment options.

One of the reason why the multidrug-resistant Gram-negative bacteriacauses serious problem is that most of the strains show resistance toantibiotics currently in use, leaving many strains untreatable. Thereare several reasons for the increase in resistant strains, but in caseof P. aeruginosa, the mutations in outer membrane and porin channel arethe main causes of the resistance. Due to these mutations, many β-lactaminhibitors are not able to enter into Gram-negative bacteria. Toovercome these resistances caused by the mutations in outer membrane andin porin channel, siderophore driven antibiotic was being heavilyresearched. Iron ions are essential ingredient for the growth ofbacteria. These iron ions have high affinity to siderosphore, andbacteria produces siderophore to bind these iron ions and internalizethem into their system. Bacteria have siderophore recognizing cellmembrane receptors to bind and internalize iron ions. FIG. 1 representsbacteria's binding mechanism to siderophore and iron ions using itsmembrane receptors.

Therefore, siderophore-mimicking moiety can be attached to antibiotic,and bacteria's siderophore receptor can bind to antibiotic. Bacteriawill then internalize the antibiotic. This internalization is mucheasier then the typical porin channel mediated antibioticinternalization, and it is also immune to resistancy cause by the porinchannel mutation. FIG. 2 represents the internalization of iron ions bybiding of siderophore to bacteria's receptor.

Although, there are many research efforts went in to overcome resistancyproblem by incorporating siderophore moiety, not many had succeed thusfar. One of the reasons is that catechol is mainly used for siderophoremoiety, but it is rapidly transformed by catechol O-methyl transferase(COMT) and can no longer be bind to siderophore receptor. Many catecholmodification has been made to overcome this issue, but they oftenresulted in low efficacy and, or high toxicity. There were also dramaticvariations on the location of the siderophore moiety in antibiotic.Therefore, there are critical needs to develop more potent antimicrobialactivity against Gram-negative bacteria drug than currently existingcephalosporins. Especially there are urgent needs to developcephalosporins against P. aeruginosa and K. pneumonia resistant strains.

SUMMARY OF INVENTION

The inventors of the present invention have synthesized novelcephalosporin derivatives represented by Chemical Formula 1,particularly novel cephalosporin compounds with a siderophore group. Thepresent invention have superior antibacterial activity as compared toexisting antibiotics, more effective against gram negative bacteria, andstronger antimicrobial activity against the major resistant strains.

Accordingly, first object of the present invention is to provide novelchemical compound represented by Chemical Formula 1.

Second object of the present invention is to provide pharmaceuticalantibiotic compositions including novel cephalosporin derivatives,prodrugs thereof, solvates thereof, isomers thereof, or pharmaceuticallyacceptable salts thereof as an effective ingredient.

Third object of the present invention is to provide method of effectiveantibiotic treatment by providing pharmaceutical antibiotic compositionsand effective amount of thereof.

DETAILED DESCRIPTION OF INVENTION

Hereinafter, the embodiments of the present invention will be describedin detail. The present invention relates to novel cephalosporinderivatives represented by Chemical Formula 1, particularly novelcephalosporin compounds with a siderophore group. The present inventionalso relates to pharmaceutical antibiotic compositions comprising anovel cephalosporin derivative represented by Chemical Formula 1, aprodrug thereof, a hydrate thereof, a solvate thereof, an isomerthereof, or a pharmaceutically acceptable salt thereof as an effectiveingredient:

wherein,

X represents CR, N, or Cl-substituted carbon (C—Cl), and where R ishydrogen or C₁-C₃ alkyl;

Y represents C₁-C₂ alkyl, CH(CH₃)CO₂H, or C(CH₃)₂CO₂H is;

L represents the CH₂ or CH═CHCH₂;

R₁ represents NH₂, NHR₁₁ or NH(CH₂)_(m)NR₁₁R₁₂ is;

R₂ represents NHR₂₁, NH(CH₂)_(n)COOH, NH(CH₂)nNR₂₁R₂₂, or NHC(═O)(CH₂)_(n)NR₂₁R₂₂ is;

Here, R₁₁, and R₂₁ independently represent hydrogen, C₁-C₃ alkyl, orselected from the followings:

R₁₂ and R₂₂ each independently represent hydrogen or C₁-C₂ alkyl;

m and n each independently represent an integer of 1 to 6;

R₃ is hydrogen or NH₂.

Cephalosporin derivatives of the present invention have effectiveantibacterial activity against antibiotic resistant Gram-negativebacteria at a lower concentration. Particularly, present invention showssuperior antimicrobial activity against P. aeruginosa, A. baumannii, andK. pneumonia as compared to currently marketed cephalosporin. When thefollowing groups are attached to the position R₁₁ and R₁₂,

the efficacy increases substantially, and in particular, the followinghydroxy piridons show excellent antibacterial activities:

An example of the cephalosporin derivative represented by ChemicalFormula 1, is represented by the compounds derived from the ChemicalFormula 2.

Wherein,

X represents CR, N, or Cl-substituted carbon (C—Cl), and where Rrepresents hydrogen or C₁-C₃ alkyl;

Y represents C₁-C₂ alkyl, CH(CH₂)CO₂H, or C(CH₂)₂CO₂H;

L represents the CH₂ or CH═CHCH₂;

R₁ represents NH₂, NHR₁₁ or NH(CH₂)_(m)NR₁₁R₁₂;

R₂ represents NHR₂₁, NH(CH₂)_(n)NR₂₁R₂₂ or NHC(═O)(CH₂)_(n)NR₂₁R₂₂;

Here, R₁₁ and R₂₁ each independently hydrogen, C₁-C₃ alkyl, or selectedfrom following groups;

R₁₂ and R₂₂ each independently represent hydrogen or C₁-C₂ alkyl;

m and n each independently represent an integer of 1 to 6.

According to the present invention, more preferred examples ofcephalosporin derivatives of Chemical Formula 2 are,

X represents CR, N, or Cl-substituted carbon (C—Cl), and where Rrepresents hydrogen or C₁-C₃ alkyl;

Y represents CH(CH₃)CO₂H, or C(CH₃)₂CO₂H;

L represents the CH2 or CH═CHCH2;

R₁ represents NH₂ or NH(CH₂)_(m)NH₂;

R₂ represents NHR₂₁, NH(CH₂)_(n)NHR₂₁ or NHC(═O) (CH₂)_(n)NHR₂₁;

R₂₁ is selected from the following groups;

m and n independently represent integers of 1 to 6 of the compounds.

As used herein the term “alkyl” includes a structure of the linear andbranch types. For example, (C₁-C₆) alkyl is methyl, ethyl, propyl,isopropyl, butyl, isobutyl, tert-butyl, pentyl and hexyl and allpossible locations and isomers.

Examples of the novel cephalosporin derivatives according to the presentinvention, but not limited to, can be presented as the compounds below.

More preferred examples of the novel cephalosporin derivatives accordingto the present invention include the following compounds:

The novel cephalosporin derivatives according to the present inventioncan be prepared into prodrugs thereof, hydrates thereof, solvatesthereof, isomers thereof or pharmaceutically acceptable salts thereof inorder to improve absorption into the body or to enhance solubility.Therefore, the prodrugs thereof, hydrates thereof, solvates thereof,isomers thereof or pharmaceutically acceptable salts thereof also fallwithin the scope of the present invention.

The terms used herein will be described briefly.

The term “pharmaceutically acceptable salt” refers to a formulation of acompound that does not cause significant irritation to an organism towhich it is administered and does not abrogate the biological activityand properties of the compound. The terms “hydrate”, “solvate” and“isomer” have the same meanings as above. The pharmaceuticallyacceptable salt thereof can be non-toxic acid added salt containingpharmaceutically acceptable anion, for example, the acid-added saltsproduced by inorganic acids such as hydrochloric acid, sulfuric acid,nitric acid, phosphoric acid, hydrobromic acid and hydrolodic acid;organocarboxylic acids such as tartaric acid, formic acid, citric acid,acetic acid, trichloroacetic acid, trifluoroacetic acid, gluconic acid,benzoic acid, lactic acid, fumaric acid and maleic acid; and sulfonicacids such as methanesulfonic acid, benzenesulfonic acid,p-toluenesulfonic acid and naphthalenesulfonic acid, can be included.Also, pharmaceutically acceptable carboxylic acid salts may be obtainedby allowing the compound of the present invention with bases to formwith metal salts or alkaline earth metal salts bases such as lithiumsalt, sodium salt, potassium salt, calcium salt, and magnesium salt;salts with amino acids such as lysine, arginine, guanidine; salts withorganic bases such as dicyclohexylamine, N-methyl-D-glutamine, tris(hydroxymethyl)methylamine, diethanolamine, choline, and triethylamine.The present invention according to Chemical Formula 1 can be convertedto its salt forms by conventional methods.

The term “hydrate” refers to a compound of the present invention or asalt thereof, that further includes a stoichiometric ornon-stoichiometric amount of water bound by non-covalent intermolecularforces. The term “solvate” as used herein means a compound of theinvention or a salt thereof, that further includes a stoichiometric ornon-stoichiometric amount of a solvent bound by non-covalentintermolecular forces. Preferred solvents are volatile, non-toxic,and/or acceptable for administration to humans. The term “isomer” meansa compound of the present invention or a salt thereof, which has thesame chemical formula or molecular formula but is optically orsterically different therefrom. Amongst these isomers structural isomerlike tautomer, asymmetric carbon center R or S isomers, geometricisomers (trans, CIS), and all stereoisomers are included. The term“prodrug” refers to an agent, which is converted into the parent drug invivo. Prodrugs are often useful because, in some situations, they may beeasier to administer than the parent drug. They may, for instance, bebioavailable by oral administration whereas the parent drug is not. Theprodrug may also have improved solubility in pharmaceutical compositionsover the parent drug. Examples of the prodrug include esters of thecompounds of the present invention and pharmaceutically acceptable saltsthereof that can be hydrolyzed in vivo. A further example of the prodrugmight be a short peptide (polyamino acid) bonded to an acidic group,where the peptide is metabolized to reveal the active moiety.

Other terms included to describe the present invention can beinterpreted typically meaning in the field.

Various types of prodrug forms are known in the related art. Forexample, refer to:

a) Design of Prodrugs, edited by H. Bundgaard, (Elsevier, 1985) andMethods in Enzymology, Vol. 42, p. 309-396, edited by K. Widder, et al.(Academic press, 1985);

b) A Textbook of Drug Design and Development, edited byKrogsgaard-Larsen and H. Bundgaard, Chapter 5 “Design and Application ofProdrugs”, by H. Bundgaard p. 113-191 (1991);

c) H. Bundgaard, Advanced Drug Delivery Reviews, 8, 1-38 (1992);

d) H. Bundgaard, et al., Journal of Pharmaceutical Sciences, 77, 285(1988); and

e) N. Kakeya, et al., Chem Pharm Bull, 32, 692 (1984).

The compounds of the present invention can represent polymorphismcompounds and polymorphism compounds with antimicrobial activity.

Novel cephalosporin derivatives according to the present invention canbe prepared in various ways depending on the type of substituents. Forexample, the composition of compounds can be prepared according to themethod illustrated below. Manufacturing methods to the proposed reactionschemes are examples only, and depending on the particular substituents,the reaction schemes can be easily transformed by those skilled in theart. Thus the exemplified reaction schemes of a method for preparingcephalosporin compounds according to the present invention are notlimited thereto, and unless otherwise stated, reactions of thesubstituents expression are the same as defined in Chemical Formula 1.

The reaction scheme of the novel cephalosporin derivatives according tothe Chemical Formula 1 is shown below.

As shown in Reaction Scheme 1, pyrimidine substituted Part A of theChemical Formula 1, and the protector (P1, P2) substituted by Part B,are reacted together and the reaction is synthesized by removing theprotectors by acid.

In the Reaction Scheme 1, t-butyl, boc, or as pmb can be used as the P1and P2 protectors, although it is not limited thereto, the halogens (Cl,Br, I, etc.) can be used as the carbon substituted leaving group in thepyrimidine reaction. Examples of Y in Part B of the Reaction Scheme 1,but not limited to, are dimethyl acetyl group protected by t-butyl, andmethyl acetyl group protected by diphenyl methyl. Polar and Aproticsolvent can be used as a solvent in this reaction. Preferred example issolvent such as DMF. Amine base such as TEA or DIPEA can be used as thebases of the reaction, but more preferred reaction method is using nobases at all.

The second reaction of the above Reaction Scheme 1 is removing theprotector by using acids like FTA or HCl.

In the first reaction of Reaction Scheme 1, the following isomers (Δ-2isomer) are created as a by-product, and in order to reduce theproduction of isomers of these by-products following the reaction shownin Reaction Scheme 2 may be made.

As shown in Reaction Scheme 2, prior to reacting with pyrimidine of PartA, create sulfoxide compound by oxidating cephem compound with MCPBAfirst and then react with Part A. React the resulting product to conductthe reduction reaction with acetyl chloride (AcCl) and KI to obtain thedesired product as the major products. In Reaction Scheme 2, methylenechloride (MC) can be used as the solvent for the MCPBA reaction, but itis not limited thereto. In addition, the reagent which can be used inoxidation and reduction reactions are not limited to MCPBA and AcCl/KI,but the oxidant and reductant with similar reactions can be used.

The invention also relates to, (a) a pharmaceutical compositionscomprising a novel cephalosporin derivative represented by ChemicalFormula 1, a prodrug thereof, a hydrate thereof, a solvate thereof, anisomer thereof, or a pharmaceutically acceptable salt thereof as aneffective ingredient, and (b) a pharmaceutical antibiotic compositionscomprising a pharmaceutically acceptable carrier thereof, diluentthereof, adjuvant thereof, or any combination thereof.

The term “pharmaceutical composition” means a mixture of the compound ofthe present invention with other chemical components such as diluents orcarriers. The above pharmaceutical composition facilitatesadministration of the compound to an organism. Multiple techniques ofadministering a compound exist in the art including, but not limited to,oral, injection, aerosol, parenteral, and topical administration.Pharmaceutical compositions can also be obtained by reacting compoundswith inorganic or organic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid,p-toluenesulfonic acid, salicylic acid and the like.

As used herein, the term “therapeutically effective amount” means theamount of active ingredient effective to alleviate or remove one or moresymptoms of the disorder to be treated, or to delay clinical markers orthe initiation of symptoms of the disease to be prevented. Thus, thetherapeutically effective amount means the amount having the effect of(1) reversing the rate of progress of the disease, (2) prohibitingfurther progress of the disease and/or (3) alleviating (preferably,removing) one or more symptoms associated with the disease. Testing thecompounds in vivo and in vitro model systems can empirically determinetherapeutically effective amount for the treatment of the disease.

The term “carrier” defines a chemical compound that facilitates theincorporation of a compound into cells or tissues. For example dimethylsulfoxide (DMSO) is a commonly utilized carrier as it facilitates theuptake of many organic compounds into the cells or tissues of anorganism.

The term “diluent” defines chemical compounds diluted in water that willdissolve the compound of interest as well as stabilize the biologicallyactive form of the compound. Salts dissolved in buffered solutions areutilized as diluents in the art. One commonly used buffered solution isphosphate buffered saline because it mimics the salt conditions of humanblood. Since buffer salts can control the pH of a solution at lowconcentrations, a buffered diluent rarely modifies the biologicalactivity of a compound.

The compound used herein may be administered as the compound per se oras a pharmaceutical composition comprising the compound with otheractive ingredients in the combination therapy or with other suitablecarriers or excipients, to the human patient. Any of the formulation andadministration techniques of the compounds in this invention may be usedas suitable and as understood in the art; “Remington's PharmaceuticalSciences,” Mack Publishing Co., Easton, Pa., 18th edition, 1990.

The pharmaceutical composition of the present invention may be preparedin a manner that is itself known, e.g. by means of conventional mixing,dissolving, granulating, dragee-making, powdering, emulsifying,encapsulating, entrapping or lyophilizing processes.

Thus, pharmaceutical compositions for use in accordance with the presentinvention may be formulated in conventional manner using one or morephysiologically acceptable carriers comprising excipients andauxiliaries, which facilitate processing of the active compounds intopreparations, which can be used pharmaceutically. Proper formulation isdependent upon the route of administration chosen. Any of the well-knowntechniques, carriers, and excipients may be used as suitable and asunderstood in the art; e.g., in Remington's Pharmaceutical Sciences,above. In the present invention according to the composition of theChemical Formula 1, injectable and oral formulation may be formulatedfor such purposes.

For injection, the agents of the present invention may be formulated inaqueous solutions or lipid emulsions, preferably in physiologicallycompatible buffers such as Hanks's solution, Ringer's solution, orphysiological saline buffer. For transmucosal administration, penetrantsappropriate to the barrier to be permeated are used in the formulation.Such penetrants are generally known in the art.

For oral administration, the compounds can be formulated readily bycombining the active compounds with pharmaceutically acceptable carrierswell known in the art. Such carriers enable the compounds of the presentinvention to be formulated as tablets, pills, powders, granules,dragees, capsules, liquids, gels, syrups, slurries, suspensions and thelike, for oral ingestion by a patient to be treated. Preferably theformulations are in capsules, tablets, pills, powders, and granulesforms, and especially, capsules and tablets forms are more useful.Tablets and pills are preferable to manufacture as theintestinal-targeted dissolving formulation. Pharmaceutical preparationsfor oral use can be obtained by mixing one or more solid excipient withpharmaceutical combination of the invention, optionally grinding theresulting mixture, and processing the mixture of granules, after addingsuitable auxiliaries, if desired, to obtain tablets or dragee cores.Suitable excipients are, in particular, fillers such as sugars,including lactose, sucrose, mannitol, or sorbitol; cellulosepreparations such as, for example, maize starch, wheat starch, ricestarch, potato starch, gelatin, gum tragacanth, methyl cellulose,hydroxypropylmethyl-cellulose, sodium carboxymethylcellulose, and/orpolyvinylpyrrolidone (PVP). If desired, disintegrating agents, such asthe cross-linked polyvinyl pyrrolidone, agar, or alginic acid or a saltthereof such as sodium alginate and magnesium stearates such aslubricants and binders may be added.

Pharmaceutical preparations, which can be used orally, include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added. Furthermore, theformulations of the present invention may be coated with entericpolymers. All formulations for oral administration should be in dosagessuitable for such administration.

The compounds may be formulated for parenteral administration byinjection, e.g., by bolus injection or continuous infusion. Formulationsfor injection may be presented in unit dosage form, e.g., in ampoules orin multi-dose containers, with an added preservative. The compositionsmay take such forms as suspensions, solutions or emulsions in oily oraqueous vehicles, and may contain formulatory agents such as suspending,stabilizing and/or dispersing agents.

In addition, an active ingredient may be for examples, a dry powderform, which can be dissolved in non-pyrogenic and non-bacterial waterprior to use.

The compounds may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter or other glycerides.

Pharmaceutical compositions suitable for use in the present inventioninclude compositions where the active ingredients are contained in anamount effective to achieve its intended purpose. More specifically, atherapeutically effective amount means an amount of compound effectiveto prevent, alleviate or ameliorate symptoms of disease or prolong thesurvival of the subject being treated. Determination of atherapeutically effective amount is well within the capability of thoseskilled in the art, especially in light of the detailed disclosureprovided herein.

When formulated into unit dose, the active ingredient of composition ofthe Chemical Formula 1 is preferably administered at a dose of from 1 to1,500 mg. Depending on the conditions of patients, including age, bodyweight, sex, administration route, state of health, and severity ofdisease, the administration dose of the compound of the presentinvention is determined according to the instructions of a physician orpharmacist. Typically, the dose ranges from about 1 to 1,500 mg per oneto three times a day for an adult. For example, the compounds of thepresent invention may be intramuscularly or intravenously injected at adose of from 1 to 1,500 mg per one to three times a day to an adult. Ahigher dose may be effective from some patients.

In addition to the compounds of the present invention, thepharmaceutical compositions of the present invention may furthercomprise (i.e., formulated together with one or more known drug(s)selected from clinically useful antibacterial agents (e.g., β-lactam,macrolide, quinolone or aminoglycoside) and anti-inflammatory agent(e.g., antifungal triazole or amphotericin), or may be administered incombination with one or more the known drug(s). Further, the compoundsof the present invention may be formulated together with or administeredin combination with a bactericidal/permeability increasing protein (BPI)product or an efflux pump inhibitor, in order to increase activityagainst Gram-negative bacteria and antibiotic resistant bacteria.

The compounds of the present invention may be formulated together withor administered in combination with vitamin, e.g., vitamin B, such asvitamin B2, vitamin B6 or vitamin B12, and folic acid. Further, thecompounds of the present invention may be formulated together with oradministered in combination with a cyclooxygenase (COX) inhibitor,particularly COX-2 inhibitor.

The present invention relates to method of antibiotic treatment usingpharmaceutical antibiotic compositions comprising a novel cephalosporinderivatives presented by Chemical Formula 1, a prodrug thereof, ahydrate thereof, a solvate thereof, an isomer thereof, or apharmaceutically acceptable salt thereof as an effective ingredient.

As described above, the present invention, a novel cephalosporinderivative, possesses superior efficacy antimicrobial activity againstGram-negative bacteria such as P. aeruginosa, K. pneumonia, A. baumanniiand also against multidrug resistant Gram-negative bacteria, andespecially against the most problematic multidrug resistantGram-negative bacteria Pseudomonas aeruginosa. In addition, thesecompounds show excellent potential as drugs during the development stageby possessing excellent pharmacokinetic profile.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 presents schematic diagram of siderophore iron (Fe) and itsreceptors;

FIG. 2 presents schematic view of the transportation process of iron ionand siderophore.

EXAMPLES

Hereinafter reference will now be made in detail to various PreparationExamples, Examples, and Test Examples. While the invention will beillustrated in conjunction with the Preparation Examples, Examples, andTest Examples, it will be understood that present description is notintended to limit the invention to those Preparation Examples, Examples,and Test Examples.

The following Preparation Examples describe preparation of the compoundsin Part A and Part B of the Reaction Scheme 1.

Preparation Example 1 Compound A-I

1-1) Preparation of Compound I:

Oxalyl chloride (1.3 mL, 15 mmol) was added to a reaction chambercontaining methylene chloride (120 mL) at −78° C. and a solution ofdimethyl sulfoxide (2.45 mL, 30 mmol) dissolved in methylene chloride(20 mL) was added. The resulting solution was stirred for 10 minutes at−78° C. A solution of N-Boc-ethanolamine (2 g, 12.4 mmol) dissolved inmethylene chloride (20 mL) was slowly added and then triethylamine (8.64ml, 62 mmol) was added. The resulting solution was stirred for 30minutes at −78° C. and additional 30 minutes at room temperature, washedwith water (100 mL) and saline (100 mL). The organic layer wasdehydrated with anhydrous sodium sulfate, concentrated under reducedpressure, and applied to column chromatography (n-hex:EA=3:1˜1:1) toyield Compound I (270 mg (14%)).

¹H NMR (600 MHz, DMSO-d₆) δ=7.83 (s, 1H), 7.49 (s, 1H), 6.88 (d, J=5.4Hz, 1H), 6.36 (br, 2H), 4.81 (br, 1H), 3.13 (m, 4H), 1.39 (s, 9H)

1-2) Preparation of Compound A-I:

4,5-diaminopyrimidine hydrochloride (2.0 g, 18.1 mmol) and Compound I(3.0 g, 18.8 mmol) were dissolved in methanol (60 mL) and then aceticacid (1.0 g, 18.1 mmol) was added. The resulting solution was stirredfor 12 hours at room temperature. Sodium cyanoborochloride (2.2 g, 36.3mmol) was added. The resulting solution was stirred for 3 hours at roomtemperature, concentrated under reduced pressure, and applied to columnchromatography (MC:MeOH=50:1˜20:1) to yield Compound A-I (1.09 g (24%)).

¹H NMR (600 MHz, chloroform-d₁) δ 8.15 (s, 1H), 7.65 (s, 1H), 5.01 (br,2H), 3.47 (br, 2H), 3.22 (t, J=5.4 Hz, 2H), 1.46 (s, 9H)

Preparation Example 2 Compound A-II

2-1) Preparation of Compound II:

Kojic acid (50 g, 0.35 mol) was dissolved in N,N-dimethylformamide (900mL) and then potassium carbonate (58.4 g, 0.42 mol) and 4-methoxybenzylchloride (61.7 g, 0.39 mol) were sequentially added. The resultingsolution was stirred for 3 hours at 80° C., concentrated under reducedpressure, and slowly added to water (800 mL) to yield a solid. The solidwas washed with ether:hexane=1:1 (800 mL) to yield Compound II (90 g(98%)).

¹H NMR (600 MHz, chloroform-d₁) δ 7.51 (s, 1H), 7.32 (d, J=8.4 Hz, 2H),6.90 (d, J=8.0 Hz, 2H), 6.45 (s, 1H), 5.00 (s, 2H), 4.45 (s, 2H), 3.81(s, 3H)

2-2) Preparation of Compound III:

Compound II (50 g, 0.19 mol) and hydroxylamine hydrochloride (66.2 g,0.95 mol) were dissolved in pyridine (620 mL). The resulting solutionwas stirred for a hours at 70° C.˜75° C., concentrated under reducedpressure, and dissolved in water (350 mL). 6N HCl (pH 1˜2) was added tothe resulting solution while stirring at 0° C. to yield a solid. Thesolid was washed with ether (300 mL) to yield Compound III (15 g (30%)).

¹H NMR (600 MHz, DMSO-d₆) δ 7.96 (s, 1H), 7.38 (d, J=8.0 Hz, 2H), 6.96(d, J=8.0 Hz, 2H0, 6.86 (s, 1H), 5.54 (br, 1H), 5.03 (s, 2H), 4.45 (s,2H), 3.74 (s, 3H)

2-3) Preparation of Compound IV:

Compound III (31 g, 0.11 mol) was dissolved in N,N-dimethylformamide(350 mL) and then potassium carbonate (31 g, 0.22 mol) and4-methoxybenzyl chloride (19.3 g, 0.12 mol) were sequentially added. Theresulting solution was stirred for 15 hours at room temperature,concentrated under reduced pressure, diluted with ethyl acetate (400mL), and filtered under reduced pressure. The filtrate was washed withwater (300 mL) and saline (300 mL). The organic layer was dehydratedwith anhydrous sodium sulfate. The resultant was washed withether:hexane=1:1 (400 mL) to yield Compound IV (42 g (95%)).

¹H NMR (600 MHz, chloroform-d₁) δ 7.27˜7.21 (m, 5H), 6.99 (s, 1H), 6.90(d, J=8.0 Hz, 2H), 6.86 (d, J=8.0 Hz, 2H), 6.49 (s, 1H), 5.03 (s, 2H),4.93 (s, 2H), 4.50 (s, 2H), 3.82 (s, 3H), 3.78 (s, 3H)

2-4) Preparation of Compound V:

Compound IV (20 g, 50.3 mmol) was dissolved in methylene chloride (580mL) and then distilled water (50 mL) was added. The resulting solutionwas stirred at 0° C. 1M sodium bromide (30 mL), 1M tetrabutyl ammoniumbromide (55 mL), TEMPO (2.36 g, 15.1 mmol), sodium hydrocarbonatesaturated solution (110 mL), and sodium hypochlorite solution (120 mL,2.01 mol) were added sequentially. The resulting solution was stirredfor 1 and a half hours at the temperature changing from 0° C. to roomtemperature. 1N HCl (pH6˜7) was added. Then, t-butanol (380 mL) wasadded and 2M 2-methyl-2-butene dissolved in tetrahydrofuran (607 mL) wassubsequently added. Thereafter, a solution of sodium chloride (45.5 g,503 mmol) and sodium dihydrogen phosphate monohydrate (52 g, 377 mmol)dissolved in distilled water (170 mL) was added. The resulting solutionwas stirred for 1 hour at room temperature. The resulting solution waspoured in a filter funnel to separate an organic layer and aqueouslayer. The organic layer was washed with sodium dihydrogen phosphatesaturated solution (800 mL), dehydrated with anhydrous sodium sulfate,concentrated under reduced pressure, and applied to columnchromatography (MC:MeOH=50:1˜8:1) to yield Compound V (40 g (61%)).

¹H NMR (600 MHz, chloroform-d₁) δ 7.35 (d, J=8.4 Hz, 2H), 7.25 (d, 8.4Hz, 2H), 6.86 (m, 4H), 6.72 (s, 1H), 6.38 (s, 1H), 6.49 (s, 1H), 5.30(s, 2H), 4.85 (s, 2H), 3.80 (s, 3H), 3.79 (s, 3H), 3.28 (m, 8H), 1.65(m, 8H), 1.42 (m, 8H), 0.99 (t, J=6.6 Hz, 12H)

2-5) Preparation of Compound A-II:

Compound VI (1.89 g, 10 mmol) was dissolved in N,N-dimethylformamide (50ml), diisopropyl ethylamine (7.2 mL, 40 mmol) and Compound V (6.52 g, 10mmol) were sequentially added, and benzotriazol-1-yl-oxytripyrrolidinophosphononium hexafluoro phosphate (6.24 g, 12 mmol) was added. Theresulting solution was stirred for 30 minutes at room temperature,diluted with ethyl acetate (300 mL), washed with water (200 mL) andsaline (150 mL), dehydrated with anhydrous sodium sulfate, concentratedunder reduced pressure, and applied to column chromatography(MC:MeOH=40:1˜10:1) to yield Compound A-II (2.2 g (40%)).

¹H NMR (600 MHz, CD₃OD) δ 7.92 (s, 1H), 7.45 (d, J=8.4 Hz, 2H), 7.35 (s,1H), 7.02 (d, J=12.6 Hz, 2H), 6.95 (d, J=12.6 Hz, 2H), 6.66 (d, J=13.2Hz, 2H), 6.41 (s, 1H), 5.33 (s, 2H), 4.77 (s, 2H), 3.79 (s, 3H), 3.73(s, 3H), 3.56 (t, J=9.0 Hz, 2H), 3.10 (t, J=9.0 Hz, 2H),

Preparation Example 3 Compound A-III

3-1) Preparation of Compound VII:

Compound II (1.0 g, 3.81 mmol) is added to 33% methylamine dissolved inethanol (19 mL). The resulting solution was stirred for 20 hours at roomtemperature, creating a white solid. The resulting solution was filteredunder reduced pressure to obtain the white solid. The white solid waswashed with ethanol (50 mL) and ether (20 mL) to yield Compound VII (778mg (75%)).

¹H NMR (600 MHz, DMSO-d₆) δ 7.53 (s, 1H), 7.34 (d, J=9.0 Hz, 2H), 6.94(d, J=9.0 Hz, 2H), 6.21 (s, 1H), 5.55 (brs, 1H), 4.91 (s, 2H), 4.36 (s,2H), 3.75 (s, 3H), 3.58 (s, 3H)

3-2) Preparation of Compound VIII:

Compound VII (778 mg, 2.83 mmol) was dissolved in dimethyl sulfoxide (7mL) and trimethylamine (1.3 g, 12.7 mmol), methylene chloride (7 mL),sulfur trioxide complex (1.35 g, 8.48 mmol) were added. The resultingsolution was stirred for 2 hours at room temperature, diluted withchloroform (150 mL), washed with water (30 mL), dehydrated withanhydrous sodium sulfate, concentrated under reduced pressure, andapplied to column chromatography (MC:MeOH=30:1˜10:1) to yield CompoundVIII (718 mg (93%)).

¹H NMR (600 MHz, chloroform-d₁) δ 9.61 (s, 1H), 7.34 (d, J=8.4 Hz, 2H),6.99 (s, 1H), 6.97 (s, 1H), 6.88 (d, J=8.4 Hz, 2H), 5.18 (s, 2H), 3.86(s, 3H), 3.80 (s, 3H)

3-3) Preparation of Compound IX:

Compound VIII (718 mg, 2.63 mmol) was dissolved in a mixture oft-butanol (8.5 mL) and tetrahydrofuran (8.5 mL) and then 2M2-methyl-2-butene (3.3 mL) dissolved in tetrahydrofuran was added. Theresulting solution was stirred at room temperature. To the resultingsolution, a solution of sodium chloride (1.9 g, 21.0 mmol) and sodiumdihydrogen phosphate monohydrate (2.1 g, 15.2 mmol) dissolved in water(8.5 mL) was added. The resulting solution was stirred for 1 hour atroom temperature, creating a white solid. The resulting solution wasfiltered under reduced pressure to obtain the white solid. The whitesolid was dissolved in water (4 mL). 1N HCl (pH 1˜2) was added. Thethus-obtained solid was filtered under reduced pressure and washed withethyl acetate (50 mL) and ether (50 mL) to yield Compound IX (510 mg(67%)).

¹H NMR (600 MHz, DMSO-d₆) δ 7.79 (s, 1H), 7.37 (d, J=12.6 Hz, 2H), 6.96(d, J=12.6 Hz, 2H), 6.71 (s, 1H), 4.97 (s, 2H), 3.83 (s, 3H), 3.76 (s,3H),

3-4) Preparation of Compound A-III:

Compound A-III (19.mg (25%)) was prepared by a method similar toPreparation Example 2-5 by using Compound VI (34.5 mg, 0.18 mmol).

¹H NMR (600 MHz, CD₃OD) δ 7.96 (s, 1H), 7.64 (s, 1H), 7.59 (s, 1H), 7.38(d, J=8.4 Hz, 2H), 6.91 (d, J=8.4 Hz, 2H), 6.56 (s, 1H), 5.02 (s, 2H),3.79 (s, 3H), 3.77 (s, 3H), 3.61 (t, J=6.0 Hz, 3.39 (t, J=6.6 Hz, 2H)

Preparation Example 4 Compound XI

4-1) Preparation of Compound X:

2-aminoethanol (2.0 g, 32.7 mmol) was dissolved in methylene chloride(110 mL) and benzyloxycarbonyl chloride (5.07 g, 29.8 mmol) andtriethylamine (4.44 g, 44.6 mmol) were sequentially added. The resultingsolution was stirred for 1 hour at room temperature. Water (40 mL) wasadded to the resulting solution. The resultant was dehydrated withanhydrous sodium sulfate and concentrated under reduced pressure toyield Compound X (4.05 g (70%)).

¹H NMR (600 MHz, chloroform-d₁) δ=7.40 (m, 5H), 5.19 (brs, 1H), 5.11 (s,2H), 3.73 (t, J=4.2 Hz, 2H), 3.37 (q, J=5.4 Hz, 2H), 2.23 (brs, 1H)

4-2) Preparation of Compound XI:

Compound XI (2.72 g (91%)) was prepared by a method similar toPreparation Example 3-2 by using Compound X (3 g, 15.3 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=9.66 (s, 1H), 7.39 (m, 5H), 5.44 (brs,1H), 5.13 (s, 2H), 4.16 (d, J=4.8 Hz, 2H)

Preparation Example 5 Compound A-IV

5-1) Preparation of Compound XII:

2,4-dichloro-5-nitropyrimidine (3 g, 15.4 mmol) was dissolved intetrahydrofuran (50 mL) and isopropyl ethylamine (2.0 g, 15.4 mmol) wasadded. To the resulting solution, N-Boc-ethyldiamine (2.48 g, 15.4 mmol)dissolved in tetrahydrofuran (20 mL) was slowly added at −78° C. whilestirring for 50 minutes and then the resulting solution was stirred for10 minutes at room temperature. The resultant was concentrated underreduced pressure and applied to column chromatography (EA:Hex=1:4˜1:3)to yield Compound XII (3.16 g (64%)).

¹H NMR (600 MHz, CDCl₃) δ 9.05 (s, 1H), 8.80 (br, 1H), 4.84 (br, 1H),3.78 (q, J=6 Hz, 2H), 3.48 (q, J=6 Hz, 2H), 1.43 (s, 9H)

5-2) Preparation of Compound XIII:

Compound XII (3.1 g, 9.75 mmol) was dissolved in methanol (50 ml) and10% palladium charcoal (1 g, 0.98 mmol) was added. The resultingsolution was applied to hydrogen purge, stirred for 40 minutes at roomtemperature, filtered with celite, and concentrated under reducedpressure to yield Compound XIII (2.8 g (99%)).

¹H NMR (600 MHz, DMSO-d⁶) δ 8.62 (br, 1H), 8.33 (s, 1H), 7.48 (s, 1H),6.99 (brs, 1H), 5.88 (brs, 2H), 3.45 (br, 2H), 3.19 (br, 2H), 1.35 (s,9H)

5-3) Preparation of Compound XIV:

Compound XIII (1.02 g, 3.52 mmol) was dissolved in 1,2-dichloroethane(34 mL) and diisopropyl ethylamine (455 mg, 3.52 mmol), Compound XI (796mg, 4.12 mmol), and sodium triacetoxy borohydride (1.12 g, 5.28 mmol)were sequentially added. The resulting solution was stirred for 3 hoursat room temperature, diluted with methylene chloride (180 mL), washedwith water (100 mL) and saline (100 mL), dehydrated with anhydroussodium sulfate, concentrated under reduced pressure, and applied tocolumn chromatography (MC:MeOH=60:1˜20:1) to yield Compound XIV (218 mg(14%)).

¹H NMR (600 MHz, CDCl₃) δ 8.22 (s, 1H), 7.60 (s, 1H), 7.34˜7.30 (m, 5H),5.77 (br, 1H), 5.46 (br, 1H), 5.18 (br, 1H), 5.11 (s, 2H), 3.58 (br,2H), 3.54 (br, 2H), 3.38 (br, 2H), 3.20 (br, 2H), 1.39 (s, 9H)

5-4) Preparation of Compound XV:

Compound XV (150 mg (100%) was prepared by a method similar toPreparation Example 5-2 by using Compound XIV (218 mg, 0.51 mmol) andwas used for next step without performing purification.

5-5) Preparation of Compound A-IV:

Compound A-IV (198 mg (57%) was prepared by a method similar toPreparation Example 5-2 by using Compound XV (150 mg, 0.51 mmol) andCompound V (330 mg, 0.51 mmol).

¹H NMR (600 MHz, CDCl₃) δ 8.22 (s, 1H), 8.44 (br, 1H), 8.09 (s, 1H),7.55 (d, J=7.8 Hz, 2H), 7.29 (s, 1H), 7.03 (s, 2H), 6.94 (d, J=8.4 Hz,2H), 6.67 (d, J=7.2 Hz, 2H), 6.44 (m, 3H), 5.49 (br, 2H), 4.48 (s, 2H),4.32 (br, 1H), 3.79 (s, 3H), 3.75 (s, 3H), 3.60 (br, 2H), 3.42 (br, 2H),3.09 (br, 2H), 2.79 (br, 2H), 1.39 (9H)

Preparation Example 6 Compound A-V

6-1) Preparation of Compound XVI:

4M HCl dissolved in 1,4-dioxane was added to Compound XIII (90 mg, 0.35mmol). The resulting solution was stirred for 1 hour at roomtemperature, distilled under reduced pressure, and dried to yieldCompound XVI (70 mg (100%)), which was used for next step withoutperforming purification.

6-2) Preparation of Compound A-V:

Compound A-V (103 mg (59%)) was prepared by a method similar toPreparation Example 2-5 by using Compound XVI (60 mg, 0.31 mmol) andCompound V (206 mg, 0.31 mmol).

¹H NMR (600 MHz, CDCl₃) δ 8.55 (br, 1H), 8.05 (s, 1H), 7.48 (s, 1H),7.40 (d, J=8.4 Hz, 2H), 7.11 (s, 1H), 6.92 (m, 4H), 6.69 (br, 1H), 6.60(d, J=7.8 Hz, 2H), 6.33 (s, 1H), 5.35 (s, 2H), 4.48 (s, 2H), 3.80 (s,3H), 3.73 (s, 3H), 3.44 (br, 2H), 3.34 (br, 2H)

Preparation Example 7 Compound A-VI

7-1) Preparation of Compound XVII:

Tert-butyl 4-hydroxybutyl carbamate (1.5 g, 7.93 mmol) was dissolved inmethylene chloride (36 mL) and imidazole (1.35 g, 19.8 mmol) andtert-butyl dimethylsilyl chloride (1.43 g, 9.51 mmol) were sequentiallyadded at 0° C. The resulting solution was diluted with ether (250 mL),washed with water (40 mL×2) and saline (40 mL×2), dehydrated withanhydrous sodium sulfate, and concentrated under reduced pressure toyield Compound XVII (2.4 g (100%)).

¹H NMR (600 MHz, CDCl₃) δ 4.68 (s, 1H), 3.63 (m, 2H), 3.13 (br, 2H),1.54 (m, 4H), 1.43 (s, 9H), 0.88 (s, 9H), 0.04 (s, 6H)

7-2) Preparation of Compound XVIII:

Compound XVII (2.5 g, 8.23 mmol) was dissolved in tetrahydrofuran (50mL) and 1.6 M n-butyl lithium dissolved in hexane was added at 0° C.Di-tert-butyl dicarbonate (2.15 g, 9.88 mmol) was then added. Theresulting solution was stirred for 2 hours at room temperature, dilutedwith ether (300 mL), washed with water (30 mL) and saline (30 mL),dehydrated with anhydrous sodium sulfate, and concentrated under reducedpressure. The resultant was dissolved in tetrahydrofuran (18 mL) and1.0M tetrabutyl ammonium bromide (14.8 mL, 14.8 mmol) dissolved intetrahydrofuran was slowly added. The resulting solution was stirred for4 and a half hours at room temperature, diluted with ether (150 mL),washed with water (30 mL×2) and saline (40 mL), dehydrated withanhydrous sodium sulfate, concentrated under reduced pressure, andapplied to column chromatography (EA:Hex=1:4) to yield Compound XVIII(1.33 g (56%)).

¹H NMR (600 MHz, CDCl₃) δ 3.68 (m, 2H), 3.61 (t, J=7.2 Hz, 2H), 1.68 (m,2H), 1.59 (m, 2H), 1.50 (s, 18H)

7-3) Preparation of Compound XIX:

Compound XVIII (330 mg, 1.14 mmol) was dissolved in dimethyl sulfoxide(2.5 mL) and diisopropyl ethylamine (300 mg, 2.30 mmol), methylenechloride (2.5 mL), and sulfur trioxide complex (370 mg, 2.28 mmol) wereadded at −20° C. The resulting solution was stirred for 30 minutes atroom temperature, diluted with ethyl acetate (150 mL), washed with water(40 mL) and saline (40 mL), dehydrated with anhydrous sodium sulfate,and concentrated under reduced pressure to yield Compound XIX (333 mg(100%)).

¹H NMR (600 MHz, CDCl₃) δ 9.78 (s, 1H), 3.63 (t, J=7.2 Hz, 2H), 2.48 (t,J=7.2 Hz, 2H), 1.91 (m, 2H), 1.50 (s, 18H)

7-4) Preparation of Compound XX:

Compound XX (220 mg (35%)) was prepared by a method similar toPreparation Example 5-3 by using 4,5-diamino pyrimidine (182 mg, 1.65mmol) and Compound XIX (948 mg, 3.30 mmol).

¹H NMR (600 MHz, CDCl₃) δ 8.09 (s, 1H), 7.63 (s, 1H), 5.84 (br, 2H),3.65 (t, J=6.6 Hz, 2H), 3.15 (t, J=6 Hz, 2H), 1.74 (m, 2H), 1.68 (m,2H), 1.50 (s, 18H)

7-5) Preparation of Compound XXI:

Compound XXI (111 mg (89%)) was prepared by a method similar toPreparation Example 6-1 by using Compound XX (220 mg, 0.58 mmol).

7-6) Preparation of Compound A-VI:

Compound A-VI (153 mg (53%)) was obtained by a method similar toPreparation Example 2-5 by using Compound XXI (110 mg, 0.50 mmol) andCompound V (330 mg, 0.50 mmol).

¹H NMR (600 MHz, CDCl₃) δ 8.36 (br, 1H), 7.98 (s, 1H), 7.44 (d, J=12.6Hz, 2H), 7.33 (s, 1H), 7.11 (br, 1H), 6.94 (m, 4H), 6.65 (d, J=12.6 Hz,2H), 6.25 (s, 1H), 5.85 (br, 2H), 5.34 (s, 2H), 4.49 (s, 2H), 3.77 (s,3H), 3.75 (s, 3H), 3.26 (br, 2H), 2.78 (br, 2H), 1.63 (br, 2H), 1.44(br, 2H)

Preparation Example 8 Compound A-VII

8-1) Preparation of Compound XXII:

Compound IV (1.0 g, 2.51 mmol) was dissolved in acetonitrile (13 mL).Manganese (IV) oxide (5.5 g, 63.5 mmol) was added at 50° C. Theresulting solution was stirred for 7 hours, filtered with celite,concentrated under reduced pressure, and applied to columnchromatography (MC:MeOH=40:1) to yield Compound XXII (0.73 g (24%)).

8-2) Preparation of Compound XXIII:

Compound VI (60 mg, 0.39 mmol) and Compound XXII (232 mg, 0.59 mmol)were dissolved in methanol (13 ml) and 10 drops of acetic acid wereadded. The resulting solution was stirred for 12 hours at roomtemperature. Sodium cyanoborohydride (370 mg, 587 mmol) was added. Theresulting solution was stirred for 3 hours at room temperature,concentrated under reduced pressure, and applied to columnchromatography (MC:MeOH=50:1˜20:1) to yield Compound XXIII (40 mg(20%)).

¹H NMR (600 MHz, CD₃OD) δ 8.05 (s, 1H), 7.50 (s, 1H), 7.20 (d, J=8.4 Hz,2H), 7.16 (d, J=8.4 Hz, 2H), 7.05 (s, 1H), 6.88 (d, J=8.4 Hz, 2H), 6.81(d, J=8.4 Hz, 2H), 6.42 (s, 1H), 5.05 (s, 2H), 4.76 (s, 2H), 3.80 (s,3H), 3.76 (s, 3H), 3.61 (s, 2H), 3.12 (br, 2H), 2.90 (br, 2H)

8-3) Preparation of Compound A-VII:

Compound XXIII (40 mg, 0.07 mmol) was dissolved in tetrahydrofuran (1mL) and methanol (0.5 mL). Di-tert-butyl dicarbonate (18 mg, 0.08 mmol)was added. The resulting solution was stirred with reflux for 1 and ahalf hours, concentrated under reduced pressure, and applied to columnchromatography (MC:MeOH=50:1˜8:1) to yield Compound A-VII (16 mg (34%)).

¹H NMR (600 MHz, CDCl₃) δ 8.08 (s, 1H), 7.49˜7.43 (br, 1H), 7.22 (Br,2H), 7.14 (br, 2H), 7.09˜7.02 (br, 1H), 6.91˜6.87 (br, 2H), 6.79 (br,2H), 6.12˜6.02 (s, 1H), 5.03 (s, 1H), 4.97 (s, 1H), 4.87 (s, 1H), 4.72(s, 1H), 4.45 (s, 1H), 4.22 (s, 1H), 3.79 (s, 3H), 3.77 (s, 3H), 3.36(br, 2H), 3.1˜3.02 (2H), 1.44˜1.35 (br, 9H)

Preparation Example 9 Compound A-VIII

9-1) Preparation of Compound A-VIII:

Compound II (2 g, 7.63 mmol) was dissolved in acetone (100 mL) withheating and Jones reagent (H₂SO₄ 1.88 mL, distilled water 6 mL, CrO₃2.14 g) was slowly added at 0° C. The resulting solution was stirred for1 hour at 0° C. and then further stirred for 1 hour at room temperature.Methanol (20 ml) was added and the resulting solution was stirred for 5minutes at room temperature. The resulting solid was removed byfiltering under reduced pressure and the filtrate was concentrated underreduced pressure. The resulting solid was washed with methanol to yieldCompound XXV (560 mg (27%)).

¹H NMR (600 MHz, DMSO-d6) δ 8.34 (s, 1H), 7.37 (d, J=11.4 Hz, 2H), 6.97(d, J=11.4 Hz, 2H), 6.92 (s, 1H), 4.90 (s, 2H), 3.76 (s, 3H),

9-2) Preparation of Compound XXVI:

Compound XXV (550 mg, 2 mmol) was dissolved in ammonia (15 mL) and theresulting solution was stirred for 2 hours with reflux. Ammonia (7 mL)was then added. The resulting solution was stirred for 1 hour withreflux, cooled to room temperature, and concentrated under reducedpressure to remove excessive ammonia. The resulting solution wasacidified with 5N HCl solution and the resulting solid was filteredunder reduced pressure to yield Compound XXVI (500 mg (91%)).

¹H NMR (600 MHz, DMSO-d6) δ 7.88 (br, 1H), 7.38 (d, J=8.4 Hz, 2H), 7.19(br, 1H), 6.91 (d, J=7.8 Hz, 2H), 5.15 (s, 2H), 3.72 (s, 3H),

9-3) Preparation of Compound XXVII:

Compound XXVI (200 mg, 0.73 mmol) was dissolved in N,N-dimethylformamide(9 mL). Potassium carbonate (1 g, 7.3 mmol) and 4-methoxybenzyl chloride(570 mg, 3.64 mmol) were sequentially added. The resulting solution wasstirred for 18 hours at 60° C., diluted with ethyl acetate (60 mL), andfiltered under reduced pressure. The filtrate was washed with water (30mL×3) and saline (30 mL). The organic layer was dehydrated withanhydrous sodium sulfate. The resultant was applied to columnchromatography (SiO₂, n-hex:EA=3:1˜1:1) to yield Compound XXVII (220 mg(59%)).

¹H NMR (600 MHz, chloroform-d₁) δ 8.22 (s, 1H), 7.70 (s, 1H), 7.39˜7.23(m, 6H), 6.88˜6.84 (m, 6H), 5.31 (s, 2H), 5.15 (s, 2H), 5.12 (s, 2H),3.79 (s, 3H), 3.78 (s, 3H), 3.77 (s, 3H)

9-4) Preparation of Compound XXVIII:

Compound XXVII (220 mg, 427 umol) was dissolved in tetrahydrofuran (11mL) and 2N potassium hydroxide aqueous solution (4.4 mL) was added. Theresulting solution was stirred for 1 and a half hours with reflux,cooled to room temperature, concentrated under reduced pressure toremove organic solvent, and acidified with 1N HCl solution. Theresulting solid was filtered under reduced pressure to yield CompoundXXVIII (160 mg (95%)).

¹H NMR (600 MHz, DMSO-d6) δ 8.31 (s, 1H), 7.74 (s, 1H), 7.35 (m, 4H),6.93 (m, 4H), 5.21 (s, 2H), 5.18 (s, 2H), 3.71 (s, 3H), 3.70 (s, 3H)

9-5) Preparation of Compound A-VIII:

Compound A-VIII (110 mg (55%) was prepared by a method similar toPreparation Example 2-5 by using Compound VI (86 mg, 455 umol) andCompound XXVIII (150 mg, 380 umol).

¹H NMR (600 MHz, DMSO-d6) δ 8.79 (t, J=5.4 Hz, 1H), 8.25 (s, 1H), 7.84(s, 1H), 7.73 (s, 1H), 7.56 (s, 1H), 7.39 (m, 4H), 6.97 (m, 4H), 6.40(br, 2H), 5.22 (s, 2H), 5.20 (s, 2H), 4.97 (t, J=5.4 Hz, 1H), 3.76 (s,3H), 3.75 (s, 3H), 3.51 (q, J=6.6 Hz, 2H), 3.21 (q, J=6 Hz, 2H)

Preparation Example 10 Compound A-IX

4,5-diaminopyrimidine hydrochloride (100 mg, 0.91 mmol) and4-dimethylaminopyridine (22 mg, 0.18 mmol) were dissolved in methylenechloride (3 mL) and 3-(tert-butoxycarbonyl)propanoic acid was added.Diisopropyl carbodiimide (138 mg, 1.1 mmol) was then slowly added at 0°C. The resulting solution was stirred at room temperature, concentratedunder reduced pressure, and applied to column chromatography(MC:MeOH=40:1˜10:1) to yield Compound A-IX (133 mg (52%)).

¹H NMR (600 MHz, chloroform-d₁) δ=8.44 (s, 1H), 8.34 (s, 1H), 8.17 (br,1H), 5.67 (br, 1H), 5.16 (br, 1H), 3.56 (q, J=9.0 Hz, 2H), 2.69 (t,J=9.0 Hz, 2H), 1.45 (s, 9H)

Preparation Example 11 Compound A-X

11-1) Preparation of Compound XXIX:

Compound XXIX (112 mg (100%)) was prepared by a method similar toPreparation Example 6-1 by using Compound A-IX (146 mg, 0.52 mmol).

11-2) Preparation of Compound A-X:

Compound A-X (102 mg (61%)) was prepared by a method similar toPreparation Example 2-5 by using Compound XXIX (63 mg, 0.29 mmol) andCompound V (189 mg, 0.29 mmol).

¹H NMR (600 MHz, CDCl₃) δ 8.29 (s, 1H), 8.19 (s, 1H), 7.27 (br, 2H),7.09 (br, 2H), 6.97 (s, 1H), 6.87 (d, J=8.4 Hz, 2H), 6.77 (d, J=7.8 Hz,2H), 6.38 (s, 1H), 5.20 (s, 2H), 4.51 (s, 2H), 3.77 (s, 3H), 3.76 (s,3H), 3.53 (br, 2H), 2.47 (br, 2H)

Preparation Example 12 Compound A-XI

Compound A-XI (147 mg (86%)) was prepared by a method similar toPreparation Example 2-5 by using Compound XXIX (66.3 mg, 0.30 mmol) andCompound XXVIII (120 mg, 0.30 mmol).

¹H NMR (600 MHz, CDCl₃) δ 8.36 (s, 1H), 8.30 (s, 1H), 8.06 (s, 1H), 7.73(s, 1H), 7.37 (d, J=8.4 Hz, 2H), 7.31 (d, J=8.4 Hz, 2H), 6.91 (d, J=8.4Hz, 2H), 6.87 (d, J=8.4 Hz, 2H), 5.16 (s, 2H), 5.13 (s, 2H), 3.80 (s,3H), 3.79 (s, 3H), 3.75 (t, J=5.4 Hz, 2H), 2.72 (t, J=6 Hz, 2H)

Preparation Example 13 Compound A-XII

Compound A-XII (146 mg (16%)) was prepared by a method similar toPreparation Example 2-5 by using 4,5-diaminopyrimidine (200 mg, 1.81mmol) and Compound V (1.18 g, 1.81 mmol).

¹H NMR (600 MHz, DMSO-d⁶) δ 8.28 (s, 1H), 8.19 (s, 1H), 8.06 (s, 1H),7.38 (m, 4H), 6.98 (m, 4H), 5.28 (s, 2H), 4.95 (s, 2H), 3.76 (s, 3H),3.75 (s, 3H)

Preparation Example 14 Compound A-XIII

14-1) Preparation of Compound XXX:

Compound XXX (12 g (71%)) was prepared by a method similar toPreparation Example 3-2 by using tert-butyl2-hydroxyethyl(methyl)carbamate (17 g, 97 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=9.61 (s, 1H), 3.92 (s, 2H), 2.92 (s,3H), 1.46 (s, 9H)

14-2) Preparation of Compound XXXI:

Compound XXXI (180 mg (38%)) was prepared by a method similar toPreparation Example 8-2 by using 4,5-diaminopyrimidine (200 mg, 1.82mmol) and Compound XXX (536 mg, 3.09 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ 8.09 (s, 1H), 7.53 (s, 1H), 6.74 (br,2H), 3.48˜3.32 (m, 4H), 2.97 (s, 3H), 1.46 (s, 9H)

14-3) Preparation of Compound XXXII:

Compound XXXII was prepared by a method similar to Preparation Example6-1 by using Compound XXXI (180 mg, 0.67 mmol) and used for next stepwithout performing purification.

14-4) Preparation of Compound A-XIII:

Compound A-XIII (30 mg (22%)) was prepared by a method similar toPreparation Example 2-5 by using Compound XXXII (70 mg, 0.34 mmol) and3-hydroxy-4-(4-methoxybenzyloxy)benzoic acid (103 mg, 0.38 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ 8.11 (s, 1H), 7.64 (s, 1H), 7.37 (d,J=8.4 Hz, 2H), 7.03 (br, 1H), 6.93 (d, J=8.4 Hz, 2H), 6.81 (br, 1H),5.21 (m, 3H), 3.80 (s, 3H), 3.42 (br, 5H), 3.07 (br, 2H)

Preparation Example 15 Compound A-XIV

Compound A-XIV (35 mg (49%)) was prepared by a method similar toPreparation Example 2-5 by using Compound XXIX (35 mg, 0.16 mmol) and3-hydroxy-4-(4-methoxybenzyloxy)benzoic acid (49 mg, 0.18 mmol).

¹H NMR (600 MHz, DMSO-d₆, +D₂O) δ 8.22 (s, 1H), 8.16 (s, 1H), 7.41 (d,J=8.4 Hz, 2H), 7.29 (d, J=1.8 Hz, 1H), 7.26 (dd, J=1.8 Hz, 9.4 Hz, 1H),7.04 (d, J=8.4 Hz, 2H), 6.95 (d, J=8.4 Hz, 2H), 5.09 (s, 2H), 3.76 (s,3H), 3.54 (t, J=7.2 Hz, 2H), 2.61 (t, J=7.2 Hz, 2H)

Preparation Example 16 Compound A-XV

Compound A-XV (221 mg (43%)) was prepared by a method similar toPreparation Example 8-2 by using Compound XXXII (200 mg, 0.98 mmol) and3,4-bis(4-methoxybenzyloxy)benzaldehyde (408 mg, 1.08 mmol).

¹H NMR (600 MHz, DMSO-d₆) δ 7.94 (s, 1H), 7.47 (s, 1H), 7.31 (m, 4H),7.08 (br, 1H), 6.99 (br d, 1H), 6.90 (m, 5H), 5.94 (br, 2H), 4.97 (s,2H), 4.94 (s, 2H), 3.71 (s, 3H), 3.70 (s, 3H), 3.30 (br, 5H), 2.85 (br,2H), 2.41 (m, 2H), 3.02 (d, J=4.2 Hz, 3H)

Preparation Example 17 Compound A-XVI

Compound A-XVI (40 mg (20%)) was prepared by a method similar toPreparation Example 2-5 by using Compound VI (95 mg, 500 umol) and3-hydroxy-4-(4-methoxybenzyloxy)benzoic acid (165 mg, 600 umol).

¹H NMR (600 MHz, DMSO-d₆) δ 9.17 (s 1H) 8.30 (t, J=6 Hz, 1H), 7.8 (s,1H) 7.52 (s, 1H), 7.36 (d, J=8.4 Hz, 2H), 7.28 (d, J=1.8 Hz, 1H), 7.24(dd, J=8.4 Hz, 2.4H, 1H), 7.00 (d, J=8.4 Hz, 1H), 6.91 (d, J=9 Hz, 2H),6.34 (br, 2H) 5.04 (s, 2H), 4.93 (br, 1H) 3.71 (s, 3H), 3.38 (q, J=6 Hz,2H), 3.17 (q, J=6 Hz, 2H)

Preparation Example 18 Compound A-XVII

Compound A-IX (450 mg, 1.6 mmol) was dissolved in anhydroustetrahydrofuran (12 mL) and lithium aluminium hydride (152 mg, 3.2 mmol)was slowly added at 0° C. The resulting solution was stirred for 3 hoursat room temperature. 15% sodium hydroxide aqueous solution (200 uL) wasthen added. The resulting solution was stirred for 1 hour at roomtemperature. The-thus obtained solid was filtrated under reducedpressure. The filtrate was concentrated under reduced pressure. Theresultant was applied to column chromatography (SiO₂, MC:MeOH=30:1˜10:1)to yield Compound A-XVII 140 mg (33%).

¹H NMR (600 MHz, chloroform-d₁+CD₃OD) δ 8.01 (s, 1H), 7.44 (s, 1H),3.23˜3.12 (m, 4H), 1.85 (m, 2H), 1.48 (s, 9H),

Preparation Example 19 Compound A-XVIII

19-1) Preparation of Compound XXXIII:

Compound XXXIII (110 mg (100%)) was prepared by a method similar toPreparation Example 6-1 by using Compound A-XVII (150 mg, 560 umol).

19-2) Preparation of Compound A-XVIII:

Compound A-XVIII (35 mg (41%)) was prepared by a method similar toPreparation Example 2-5 by using Compound XXXIII (50 mg, 245 umol) and3-hydroxy-4-(4-methoxybenzyloxy)benzoic acid (92 mg, 335 umol).

¹H NMR (600 MHz, DMSO-d₆) δ 9.15 (s, 1H), 8.24 (t, J=6 Hz, 1H), 7.78 (s,1H), 7.38 (s, 1H), 7.37 (d, J=8.4 Hz, 2H), 7.26 (d, J=2.4 Hz, 1H), 7.22(dd, J=8.4 Hz, 2.4 Hz, 1H), 6.99 (d, J=8.4 Hz, 1H), 6.90 (d, J=8.4 Hz),6.37 (br, 2H), 5.03 (s, 2H), 4.73 (t, J=5.4 Hz, 1H), 3.71 (s, 3H), 3.30(m, 2H), 3.04 (q, J=6.6 Hz, 2H), 1.80 (m, 2H)

Preparation Example 20 Compound A-XIX

20-1) Preparation of Compound XXXIV:

Compound XXXIV (854 mg (70%)) was prepared by a method similar toPreparation Example 2-5 by using 4,5-diaminopyrimidine (500 mg, 4.54mmol) and 2-(tert-butoxycarbonylamino)acetic acid (876 mg, 4.99 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ 8.46 (s, 1H), 8.25 (br, 1H), 8.11 (br,1H), 5.45 (br d, 3H), 3.91 (s, 2H), 1.48 (s, 9H)

20-2) Preparation of Compound XXXV:

Compound XXXV (650 mg (100%)) was prepared by a method similar toPreparation Example 6-1 by using Compound XXXIV (854 mg, 3.19 mmol).

20-3) Preparation of Compound A-XIX:

Compound A-XIX (357 mg (68%)) was prepared by a method similar toPreparation Example 2-5 by using Compound XXXV (250 mg, 1.23 mmol) and3-hydroxy-4-(4-methoxybenzyloxy)benzoic acid (337 mg, 1.23 mmol).

¹H NMR (600 MHz, DMSO-d₆) δ 8.24 (s, 1H), 8.14 (s, 1H), 7.41 (d, J=8.4Hz, 2H), 7.35˜7.31 (m, 2H), 7.07 (d, J=8.0 Hz, 1H), 6.95 (d, J=8.4 Hz,2H), 6.79 (br, 2H), 5.10 (s, 2H), 4.02 (d, J=6.0 Hz, 2H)

Preparation Example 21 Compound A-XX

21-1) Preparation of Compound XXXVI:

4-bromoasophthalic acid (1 g, 4.1 mmol) was dissolved in distilled water(4 mL) and sodium bicarbonate (1.2 g, 11 mmol) was added. The resultingsolution was stirred for 1 and a half our at 85° C. N¹, N¹, N²,N²-tetramethylethane-1,2-diamine (31 mg, 270 umol) and cooper bromide(18 mg, 126 umol) were dissolved in distilled water (0.5 mL). Theresulting solution was stirred for 1 hour. The two resulting solutionswere mixed and stirred for 18 hours at 85° C., cooled to roomtemperature, and acidified with 1N HCl aqueous solution. The resultingsolid was filtered under reduced pressure, washed with water, and driedunder reduced pressure to yield Compound XXXVI (720 mg (97%)).

¹H NMR (600 MHz, DMSO-d₆) δ 8.34 (t, J=3.6 Hz, 1H), 8.04 (dt, J=1.2 Hz,13.2 Hz, 1H), 7.05 (dd, J=3.6 Hz, 12.6 Hz, 1H)

21-2) Preparation of Compound XXXVII:

Compound XXXVI (720 mg, 3.95 mmol) was dissolved in trifluoroacetic acid(4.32 mL). Acetone (2 mL) and trifluoroacetic anhydride (TFAA) (1.45 mL)were added sequentially. The resulting solution was stirred for 8 hoursat 100° C., cooled to room temperature, and concentrated under reducedpressure. The resultant was dissolved in ethyl acetate (100 mL) andwashed with 1N HCl aqueous solution (50 mL). The organic layer wasdehydrated with anhydrous sodium sulfate and concentrated under reducedpressure. The thus-obtained solid was filtered under reduced pressure toyield Compound XXXVII (500 mg (57%)).

¹H NMR (600 MHz, chloroform-d₁) δ 8.76 (d, J=2.4 Hz, 1H), 8.30 (dd,J=9.0 Hz, 1.8 Hz, 1H), 7.08 (d, J=8.4 Hz, 1H), 1.78 (s, 6H)

21-3) Preparation of Compound A-XX:

Compound A-XX (90 mg (72%)) was prepared by a method similar toPreparation Example 2-5 by using Compound VI (80 mg, 416 umol) andCompound XXXVII (78 mg, 350 umol).

¹H NMR (600 MHz, CD₃OD) δ 8.42 (d, J=3.6 Hz, 1H), 8.13 (m, 1H), 7.94 (s,1H), 7.61 (s, 1H), 7.14 (d, 12.6 Hz, 1H) 3.66 (t, 9.6 Hz, 2 Hz) 3.40 (t,9.6 Hz, 2H), 1.74 (s, 6H),

Preparation Example 22 Compound A-XXI

22-1) Preparation of Compound XXXVIII:

Ethyl 3,4-dihydroxybenzoate (5 g, 28 mmol) was dissolved inN,N-dimethylformamide (50 mL) and potassium carbonate (15 g, 110 mmol)was added. The resulting solution was stirred for 2 days at roomtemperature, diluted with ethyl acetate (400 mL), and filtered underreduced pressure. The filtrate was washed with water (300 mL×3) andsaline (300 mL). The organic layer was concentrated under reducedpressure. Hexane was added to the resultant. The thus-obtained solid wasfiltered under reduced pressure to yield Compound XXXVIII (11 g (97%)).

¹H NMR (600 MHz, chloroform-d₁) δ 7.64 (d, J=1.8 Hz, 1H), 7.63 (dd,J=7.8 Hz, 2.4 Hz, 1H), 7.38 (m, 4H), 6.93 (m, 5H), 5.13 (s, 2H), 5.11(s, 2H), 4.35 (q, J=7.2 Hz, 2H), 3.81 (s, 6H), 1.38 (t, J=7.2 Hz, 3H)

22-2) Preparation of Compound XXXIX:

Compound XXXVIII (11 g, 27 mmol) was dissolved in tetrahydrofuran (120mL) and ethanol (130 mL). 2N lithium hydroxide aqueous solution (52 mL)was added. The resulting solution was stirred for 12 hours at roomtemperature, concentrated under reduced pressure, diluted with distilledwater (200 mL), and washed with ethyl acetate (200 mL). Thethus-obtained aqueous solution layer was acidified with 1N HCl aqueoussolution. The thus-obtained solid was filtered under reduced pressureand vacuum dried to yield Compound XXXIX (8.6 g (81%)).

¹H NMR (600 MHz, DMSO-d₆) δ 7.50 (m, 2H), 7.34 (m, 4H), 7.11 (d, J=9.6Hz, 1H), 6.91 (m, 4H), 5.07 (s, 2H), 5.02 (s, 2H), 3.71 (s, 6H)

22-3) Preparation of Compound A-XXI:

Compound A-XXI (300 mg (57%)) was prepared by a method similar toPreparation Example 2-5 by using Compound VI (190 mg, 1 mmol) andCompound XXXIX (410 mg, 1.1 mmol).

¹H NMR (600 MHz, DMSO-d₆) δ 8.46 (t, J=5.4 Hz, 1H), 7.84 (s, 1H), 7.56(d, 11.4 Hz, 2H), 7.46 (dd, J=9.0 Hz, 2.4 Hz, 1H), 7.37 (d, J=9 Hz, 4H),7.132 (d, J=8.4 Hz, 1H), 6.94 (d, J=9 hz, 4H), 6.37 (br, 2H) 5.09 (s,2H), 5.06 (s, 2H), 4.99 (t, J=5.4 Hz, 1H), 3.75 (s, 6H), 3.45 (q, J=6Hz, 2H), 3.26 (q, J=6 Hz, 2H)

Preparation Example 23 Compound A-XXII

Compound A-XXII (135 mg (59%)) was prepared by a method similar toPreparation Example 8-2 by using 4,5-diaminopyrimidine hydrochloride(100 mg, 0.91 mmol) and tert-butyl 4-oxobutanoate (158 mg, 0.99 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=8.11 (s, 1H), 7.33 (s, 1H), 6.43 (br,2H), 5.06 (br, 1H), 3.16 (t, J=6.6 Hz, 2H), 2.48 (t, J=6.6 Hz, 2H), 2.02(m, 2H), 1.47 (s, 9H)

Preparation Example 24 Compound A-XXIII

24-1) Preparation of Compound XL:

2,4-dichloro-5-nitropyrimidine (3 g, 15.4 mmol) was dissolved intetrahydrofuran (52 mL) and 2N methylamine (15.4 mL) dissolved intetrahydrofuran was slowly added at −78° C. The resulting solution wasstirred for 10 minutes and then further stirred for 50 minutes at roomtemperature. The resulting solution was concentrated under reducedpressure, diluted with ethyl acetate (50 mL), and washed with water (30mL) and saline (30 mL). The resultant was dehydrated with anhydroussodium sulfate, concentrated under reduced pressure, and applied tocolumn chromatography (EA:Hex=20:1˜5:1) to yield Compound XL (925 mg(32%)).

¹H NMR (600 MHz, chloroform-d₁) δ 9.05 (s, 1H), 8.41 (br, 1H), 3.23 (d,J=4.8 Hz, 3H),

24-2) Preparation of Compound A-XXIII:

Compound XL (506 mg, 2.68 mmol) was dissolved in methanol (5 mL) and 10%palladium charcoal (285 mg, 0.27 mmol) was added. The resulting solutionwas applied to hydrogen gas purge, stirred for 2 hours at roomtemperature, filtered with celite, and concentrated under reducedpressure to yield Compound A-XXIII (411 mg (98%)).

¹H NMR (600 MHz, DMSO-d₆) δ 8.87 (br, 1H), 8.37 (s, 1H), 7.47 (s, 1H),5.94 (br, 2H), 3.02 (d, J=4.2 Hz, 3H)

Preparation Example 25 Compound A-XXIV

Compound VI (50 mg, 0.32 mmol) was dissolved in methanol (1 mL).Triethylamine (40 mg, 0.40 mmol) and tert-butylimino(1H-pyrazole-1-yl)methylcarbamate (69 mg, 0.32 mmol) were added.The resulting solution was stirred for 15 hours at room temperature,concentrated under reduced pressure, and applied to columnchromatography (MC:MeOH=10:1˜7:1) to yield Compound A-XXIV (24 mg(25%)).

¹H NMR (600 MHz, CD₃OD) δ 8.02 (br, 1H), 7.60 (br, 1H), 3.64 (t, J=5.4Hz, 2H), 3.44 (br, 2H), 1.55sm, 9H)

Preparation Example 26 Compound A-XXV

Compound A-XXV (189 mg (50%)) was prepared by a method similar toPreparation Example 8-2 by using Compound XLI (192 mg, 0.68 mmol) andCompound XIX (333 mg, 1.16 mmol).

¹H NMR (600 MHz, CDCl₃) δ 8.19 (s, 1H), 7.49 (s, 1H), 6.07 (br, 1H),4.92 (br, 1H), 3.64 (t, J=7.2 Hz, 2H), 3.53 (br, 2H), 3.20 (br, 2H),3.11 (t, J=6.6 Hz, 2H), 1.72 (m, 6H), 1.60 (m, 2H), 1.50˜1.44 (m, 27H)

Preparation Example 27 Compound A-XXVI

Compound A-XXVI (228 mg (59%)) was prepared by a method similar toPreparation Example 2-5 by using 4,5-diaminopyrimidine (144 mg, 1.31mmol) and 4-(tert-butoxycarbonylamino)butanoic acid (266 mg, 1.31 mmol).

¹H NMR (600 MHz, CD₃OD) δ 8.30 (s, 1H), 8.24 (s, 1H), 3.18 (t, J=6 Hz,2H), 2.51 (t, J=6.6 Hz, 2H), 1.89 (t, J=6.6 Hz, 2H), 1.47 (s, 9H)

Preparation Example 28 Compound A-XXVII

28-1) Preparation of Compound XLII:

Compound XLII (180 mg (100%)) was prepared by a method similar toPreparation Example 6-1 by using Compound A-XXVI (232 mg, 0.78 mmol).

28-2) Preparation of Compound A-XXVII:

Lithium aluminium hydride (87 mg, 2.3 mmol) was added to tetrahydrofuran(5 mL) and the resulting solution was stirred at room temperature. Afteradding Compound XLII (180 mg, 0.78 mmol), the resulting solution wasstirred for 20 minutes with reflux. 15% sodium hydroxide aqueoussolution (0.1 mL) was added. The thus-obtained solid was filtered underreduced pressure. The filtrate was concentrated under reduced pressureand dissolved in methanol (4 mL). Di-tert-butyl dicarbonate (171 mg,0.78 mmol) was added. The resulting solution was stirred for 30 minuteswith reflux, concentrated under reduced pressure, and applied to columnchromatography (MC:MeOH=45:1˜15:1) to yield Compound A-XXVII (25.6 mg(11%)).

¹H NMR (600 MHz, CDCl₃) δ 8.14 (s, 1H), 7.62 (s, 1H), 3.28 (br, 2H),3.15 (br, 2H), 1.71 (m, 2H), 1.64 (m, 2H), 1.45 (s, 9H)

Preparation Example 30 Compound A-XXVIII

Compound A-XXVIII (135 mg (55%)) was prepared by a method similar toPreparation Example 2-5 by using 4,5,6-triaminopyrimidine sulfate (200mg, 0.86 mmol) and 2-(tert-butoxycarbonylamino)acetic acid (166 mg, 0.95mmol).

¹H NMR (600 MHz, CD₃OD) δ 7.83 (s, 1H), 3.82 (s, 2H), 1.46 (s, 9H)

Preparation Example 31 Compound A-XXIX

Compound A-XXIX (257 mg (62%)) was prepared by a method similar toPreparation Example 8-2 by using 4,5,6-triaminopyrimidine (193 mg, 1.54mmol) and Compound I (370 mg, 2.31 mmol).

¹H NMR (600 MHz, CD₃OD) δ 7.89 (s, 1H), 3.19 (t, J=6 Hz, 2H), 2.90 (t,J=6 Hz, 2H), 1.42 (s, 9H)

Preparation Example 32 Compound A-XXX

Compound A-XXX (112 mg (44%)) was prepared by a method similar toPreparation Example 2-5 by using 4,5,6-triaminopyrimidine sulfate (200mg, 0.86 mmol) and 2-(tert-butoxycarbonylamino)acetic acid (179 mg, 0.95mmol).

¹H NMR (600 MHz, CD₃OD) δ 7.81 (s, 1H), 3.42 (t, J=6.6 Hz, 2H), 2.60 (t,J=6 Hz, 2H), 1.43 (s, 9H)

Preparation Example 33 Compound A-XXXI

Compound A-XXXI (420 mg (60%)) was obtained as a white solid by a methodsimilar to Preparation Example 2-5 by using 4,5,6-triaminopyrimidinesulfate (500 mg, 2.24 mmol) and 3-(tert-butoxycarbonylamino)butanoicacid (500 mg, 2.46 mmol).

¹H NMR (400 MHz, DMSO d-₆) δ 8.47 (s, 1H), 7.72 (s, 1H), 6.85 (t, J=8.4Hz, 1H), 5.87 (brs, 4H), 2.94 (m, 2H), 2.30 (t, J=10.8 Hz, 2H), 1.64 (m,2H), 1.39 (s, 9H)

Preparation Example 34 Compound A-XXXII

34-1) Preparation of Compound XLIII:

Crotonic acid (4 g, 47 mmol) was dissolved in CCl₄ (30 mL). NBS (9 g, 51mmol) and benzoylperoxide (75 mg, 0.3 mmol) were added with reflux over3 hours. (i.e., o hour: 4 g, 25 mg; after 1 hour: 3 g, 25 mg; after 2hours: 2 g, 25 mg) The resulting solution was cooled to roomtemperature, diluted with methylene chloride (20 mL), and washed with0.2 N HCl aqueous solution. The organic layer was dehydrated withanhydrous sodium sulfate and concentrated under reduced pressure toyield Compound XLIII (2.2 g (27%)).

¹H NMR (600 MHz, chloroform-d₁) δ=7.10 (q, J=7.2 Hz, 1H), 6.01 (dt, J=15Hz, 1.2 Hz 1H), 4.00 (dd, J=7.2 Hz, 0.6 Hz, 2H)

34-2) Preparation of Compound XLIV:

Compound XLIII (1 g, 6.06 mmol) was dissolved in anhydrous methylenechloride (20 mL). At room temperature, oxalyl chloride (1.03 mL, 12mmol) was added and 3 drops of N,N-dimethylformamide were added. Theresulting solution was stirred for 1 hour at room temperature anddistilled under reduced pressure to remove the solvent. The resultantwas dissolved in tetrahydrofuran (12 mL) and cooled to 0° C. by usingice water. O-(4-methoxybenzyl)hydroxylamine (928 mg, 6 mmol) dissolvedin tetrahydrofuran (20 mL) was slowly added. Diisopropylethylamine (4.1mL, 21 mmol) was then added. The resulting solution was stirred for 30minutes at 0° C. and for 20 minutes at room temperature. The resultantwas concentrated under reduced pressure, diluted with ethyl acetate 30ml, and washed with water 30 mL and saline 30 mL. The organic layer wasdehydrated with anhydrous sodium sulfate and concentrated under reducedpressure. The resultant was applied to column chromatography(n-Hex:EA=3:1) to yield Compound XLIV (515 mg (28%)).

¹H NMR (600 MHz, chloroform-d₁) δ 7.30 (d J=7.8 Hz 2H) 7.00 (q, J=7.2Hz, 1H), 6.96 (d, J=7.2 Hz 2H) 4.86 (br, 2H), 4.00 (d, J=11.42 Hz, 2H),3.79 (s, 2H)

34-3) Preparation of Compound A-XXXII:

Compound XLIV (150 mg, 0.5 mmol) and 4,5-diaminopyrimidine weredissolved in N,N-dimethylformamide (1 mL). The resulting solution wasstirred for 12 hours at room temperature, diluted with ethyl acetate (10mL), and washed with water (10 ml). The organic layer was dehydratedwith anhydrous sodium sulfate and concentrated under reduced pressure.The resultant was applied to column chromatography to yield CompoundA-XXXII (5 mg (3%)).

¹H NMR (600 MHz, chloroform-d₁+CD₃OD) δ 7.98 (s, 1H) 7.30 (m 3H) 6.84(m, 3H), 5.87 (br, 1H) 4.86 (br, 2H), 3.83˜3.75 (m, 5H)

Preparation Example 35 Compound A-XXXIII

Compound A-XXXIII (854 mg (71%)) was obtained by a method similar to

Preparation Example 2-5 by using 4,5-diaminopyrimidine (500 mg, 4.54mmol) and 2-(tert-butoxycarbonylamino)acetic acid (876 mg, 4.99 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ 8.46 (s, 1H), 8.24 (s, 1H), 8.11 (s,1H) 5.49 (br, 2H), 5.41 (s, 1H), 3.91 (s, 2H), 1.47 (s, 9H)

Preparation Example 36 Compound XLV

GCLE ((6S,7R)-4-methoxybenzyl3-(chloromethyl)-8-oxo-7-(2-phenylacetamido)-5-thia-1-azabicyclo[4.2.0]oct-2-ene-2-carboxylate)(49 g, 0.1 mol) was dissolved in methylene chloride (700 mL). A solutionof pyridine (15.8 g, 0.2 mol) and phosphorous pentachloride (33.3 g,0.16 mol) dissolved in methylene chloride (350 mL) was added at 0° C.The resulting solution was stirred for 2 hours at 0° C. and cooled to−40° C. After adding methanol (80 mL), the resulting solution wasstirred for 10 minutes at −40° C. and for 2 hours at 0° C., diluted withmethylene chloride (400 mL), and washed with 5% sodium bicarbonateaqueous solution (800 mL×2) and 1N HCl aqueous solution (1 L). Theorganic layer was dehydrated with anhydrous sodium sulfate andconcentrated under reduced pressure to about 200 mL. Ether (3 L) wasadded and the thus-obtained solid was filtered under reduced pressure toyield Compound XLV (30 g (74%)).

¹H NMR (600 MHz, DMSO-d6) δ 8.8 (br, 2H), 7.33 (d, J=12.6 Hz, 2H), 6.91(d, J=12.6 Hz, 2H), 5.23 (m, 4H), 4.55 (dd, J=17.4 Hz, 57 Hz, 2H), 3.78(dd, J=26.4 Hz, 70.8 Hz, 2H), 3.71 (s, 3H)

Preparation Example 37 Compound XLVII

Ethyl 2-oxo-2-(2-(tritylamino)thiazol-4-yl)acetate (100 g, 230 mmol) wasdissolved in methanol (95 mL). A solution of sodium hydroxide (9.4 g,235 mmol) dissolved in methanol (235 mL) was added. The resultingsolution was stirred for 10 minutes with reflux. The thus-obtained solidwas filtered under reduced pressure, washed with methanol, dissolved inwater (200 mL), and acidified with 2N HCl aqueous solution. Theresulting solid was filtered under reduced pressure to yield CompoundXLVII (84 g (84%)).

¹H NMR (600 MHz, DMSO-d6) δ 9.03 (s, 1H), 7.79 (s, 1H), 7.34˜7.21 (m,15)

Preparation Example 38 Compound B-1

38-1) Preparation of Compound XLVIII:

Benzophenone (25 g, 137 mmol) was dissolved in ethanol (250 mL) andhydrazine monohydrate (13.7 g, 274 mmol) was added. The resultingsolution was stirred for one day with reflux. Then, additional hydrazinemonohydrate (13 g) was added and the resulting solution was stirred forone day with reflux. After the starting material, benzophenone,disappeared, the resulting solution was concentrated under reducedpressure to remove ethanol, diluted with ethyl acetate (500 mL), andwashed with water (300 mL×2) and saline (200 mL). The organic layer wasdehydrated with anhydrous sodium sulfate and concentrated under reducedpressure. The thus-obtained solid was filtered under reduced pressure toyield Compound XLVIII (20 g (75%)). The filtrate was recrystallized withethyl acetate and hexane to yield Compound XLVIII (4 g (15%)).

¹H NMR (400 MHz, DMSO-d6) δ 7.55˜7.17 (m, 10H), 6.20 (s, 2H)

38-2) Preparation of Compound XLIX:

Compound XLVIII (20 g, 102 mmol) was dissolved in ether (320 mL). Sodiumsulfate (Na2SO4, 22 g, 153 mmol), saturated potassium hydroxide ethanolsolution (8 mL, 40 g/100 mL EtOH), and mercury oxide (HgO, 55 g, 255mmol) were sequentially added. The resulting solution was stirred at ahigh speed for 1 hour. The thus-created solid was filtered under reducedpressure with celite and the filtrate was concentrated under reducedpressure to yield Compound XLIX (19 g (100%)), which was used for nextstep without performing purification.

¹H NMR (400 MHz, chloroform-d₁) δ 7.39˜7.15 (m, 10H),

38-3) Preparation of Compound L:

(+) lactic acid (85%, 9.6 g, 90 mmol) was dissolved in ethyl acetate(400 mL) and a solution of Compound XLIX (19 g, 100 mmol) dissolved inethyl acetate (200 mL) was added at 0° C. over 20 minutes. The resultingsolution was stirred for 12 hours at room temperature. The resultant wasconcentrated under reduced pressure and applied to column chromatography(SiO2, EA:n-hex=1:6) to yield Compound L (17 g (76%)).

[α]_(D)=−9.41 (C=5.00, CHCl₃)

¹H NMR (600 MHz, chloroform-d₁) δ=7.35˜7.24 (m, 10H), 6.92 (s, 1H), 4.39(m, 1H), 2.76 (d, J=5.4 Hz, 1H), 1.47 (d, J=7.2 Hz, 3H)

38-4) Preparation of Compound LI:

Compound L (10 g, 39 mmol) was dissolved in N,N-dimethylformamide (40mL). The resulting solution was cooled to 0° C. and sulfuryl chloride(5.79 g, 42.9 mmol) was slowly added over 15 minutes. The resultingsolution was stirred for 20 minutes at 0° C. and for 1 and a half hourat room temperature and diluted with ethyl acetate (200 ml). Cool sodiumbicarbonate solution (100 mL) was added to end the reaction. Theresulting solution was washed with saturated sodium bicarbonate solution(100 mL) and saline (100 mL). The organic layer was dehydrated withanhydrous sodium sulfate and the filtrate was concentrated under reducedpressure. The resultant was applied to column chromatography (SiO₂,EA:n-hex=1:9) to yield Compound LI (5.3 g (49%)).

¹H NMR (600 MHz, chloroform-d₁) δ=7.36˜7.26 (m, 10H), 6.90 (s, 1H), 4.51(q, J=6.6 Hz, 1H), 1.72 (d, J=6.6 Hz, 3H)

38-5) Preparation of Compound LII:

Compound LI (4 g, 14.6 mmol) was dissolved in N,N-dimethylformamide (30mL). N-hydroxy phthalamide (2.45 g, 15 mmol) and potassium carbonate(2.07 g, 15 mmol) were added. The resulting solution was stirred for 12hours at room temperature, diluted with ethyl acetate (400 mL), washedwith water (200 mL) and saline (200 mL×2), and recrystallized withhexane to yield Compound LII (6 g (99%)).

[α]_(D)=−63.26 (C=5.00, CHCl₃)

¹H NMR (600 MHz, chloroform-d₁) δ=7.76 (m, 4H), 7.32˜7.19 (m, 10H), 6.91(s, 1H), 5.05 (q, J=6.6 Hz, 1H), 1.65 (d, J=7.2 Hz, 3H)

38-6) Preparation of Compound LIII:

Compound LII (2.5 g, 6.23 mmol) was dissolved in methylene chloride (10mL). Methyl hydrazine (287 mg, 6.23 mmol) was added at 0° C. Theresulting solution was stirred for 2 hours at 0° C. to create a solid.The solid was filtered under reduced pressure and the filtrate wasconcentrated under reduced pressure to yield Compound LIII, which wasused for next step without performing purification.

38-7) Preparation of Compound LIV:

Compound LIII was dissolved in methylene chloride (5 mL) and methanol(20 mL). Compound XLVII (2.6 g, 6.2 mmol) was added at 0° C. Theresulting solution was stirred for 30 minutes at 0° C. and 3 hours atroom temperature, concentrated under reduced pressure to remove thesolvent, diluted with ethyl acetate (150 mL), and washed with 0.1N HClaqueous solution (100 mL) and saline (100 mL). The organic layer wasdehydrated with anhydrous sodium carbonate, concentrated under reducedpressure to remove the solvent, and crystallized with ethyl acetate andhexane to yield Compound LIV (2.5 g (60%)).

¹H NMR (600 MHz, DMSO-d6) δ 8.87 (s, 1H), 7.44 (m, 25H), 6.82 (s, 2H),4.89 (q, J=7.2 Hz, 1H), 1.41 (d, J=6.6 Hz, 3H)

38-8) Preparation of Compound B-1

Compound LIV (2.07 g, 4.03 mmol) and Compound XLV (1.8 g, 4.44 mmol)were dissolved in methylene chloride (45 mL). At 0° C., pyridine (1.47mL, 18 mmol) and phosphoryl chloride (POCl3, 376 uL, 4.03 mmol) wereadded. The resulting solution was stirred for 20 minutes at 0, dilutedwith ethyl acetate (150 mL), washed with water (50 mL) and saline (30mL×2). The organic layer was dehydrated with anhydrous sodium sulfate,concentrated under reduced pressure, and applied to columnchromatography (SiO₂, n-hex:EA=6:1˜2:1) to yield Compound B-1 (1.9 g(47%)).

¹H NMR (600 MHz, chloroform-d₁) δ=8.10 (d, J=8.4 Hz, 1H), 7.34˜7.22 (m,27H), 6.98 (s, 1H), 6.91 (s, 1H), 6.89 (d, J=7.8 Hz, 2H), 6.73 (s, 1H),5.91 (dd, J=4.8 Hz, 8.4 Hz, 1H), 5.26 (m, 4H), 4.94 (d, J=5.4 Hz, 1H),4.58 (dd, J=11.4 Hz, 94.8 Hz, 2H), 3.79 (s, 3H), 3.546 (dd, 18 Hz, 105.6Hz, 2H), 1.61 (d, J=6.6 Hz, 3H)

Preparation Example 39 Compound B-II

39-1) Preparation of Compound LVI:

Compound LV (30 g, 149 mmol) was dissolved in tetrahydrofuran (500 mL).Di-tert-butyl dicarbonate (33 g, 152 mmol) and 4-dimethylaminopyridine(388 mg, 3.17 mmol) were sequentially added. The resulting solution wasstirred for 20 hours at room temperature, concentrated under reducedpressure, and applied to column chromatography (EA:Hex=1:7˜1:5) to yieldCompound LVI (17.7 g (39%)).

¹H NMR (600 MHz, chloroform-d₁) δ=8.58 (br, 1H), 8.27 (s, 1H), 4.44 (q,J=7.8 Hz, 2H), 1.54 (s, 9H), 1.42 (t, J=6.6 Hz, 3H)

39-2) Preparation of Compound LVII:

Compound LVI (17.7 g, 58.9 mmol) was dissolved in methanol (118 mL) andthe resulting solution was stirred at room temperature. Sodium hydroxide(4.24 g, 106 mmol) dissolved in distilled water (40 mL) was slowlyadded. The resulting solution was stirred for 1 hour, concentrated underreduced pressure, dissolved in water (200 mL), and solidified with 1NHCl (pH 1˜2). The thus-obtained solid was filtered under reducedpressure and washed with water (200 mL) to yield Compound LVII (16 g(100%)).

¹H NMR (600 MHz, DMSO-d6) δ 11.92 (s, 1H), 8.35 (s, 1H), 1.47 (s, 9H)

39-3) Preparation of Compound LVIII:

Compound LVII (16 g, 58.7 mmol) was dissolved in 1,4-dioxane (118 mL)and N-chlorosuccineimide (NCS, 8.1 g, 60.7 mmol) was added. Theresulting solution was stirred for 30 minutes at room temperature andfor 15 hours at 40. The resulting solution was filtered under reducedpressure and the filtrate was concentrated under reduced pressure. Theresultant was filtered under reduced pressure with ether/n-hexane=2/1(150 mL) to remove the thus-created solid and the filtrate wasconcentrated under reduced pressure to yield Compound LVIII (12.3 g(68%)).

¹H NMR (600 MHz, DMSO-d6) δ 12.19 (s, 1H), 1.43 (s, 9H)

39-4) Preparation of Compound LIX:

Compound LIX (14.2 g (63%)) was prepared by a method similar toPreparation Example 38-7 by using Compound LVIII (12.3 g, 40.1 mmol) andCompound LIII (12.8 g, 47.2 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=7.34˜7.27 (m, 10H), 6.92 (s, 1H), 5.09(q, J=6.6 Hz, 1H), 1.60 (d, J=7.2 Hz, 3H), 1.51 (s, 9H)

39-5) Preparation of Compound B-II:

Compound B-II (2.62 g (54%)) was prepared by a method similar toPreparation Example 38-8 by using Compound LIX (3.0 g, 5.35 mmol) andCompound XLV (2.82 g, 6.96 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ 7.96 (br, 1H), 7.86 (d, J=9.6 Hz, 1H),7.35 (d, J=9 Hz, 2H), 7.29˜7.25 (m, 10H), 6.92˜6.87 (m, 3H), 6.03 (q,J=4.8 Hz, 1H), 5.27 (d, J=11.4 Hz, 1H), 5.21 (d, J=11.4 Hz, 1H), 5.10(q, J=7.2 Hz, 1H), 4.98 (d, J=5.4 Hz, 1H), 4.60 (d, J=12 Hz, 1H), 4.44(d, J=12 Hz, 1H), 3.81 (s, 3H), 3.59 (d, J=18.6 Hz, 1H), 3.41 (d, J=18Hz, 1H), 1.64 (d, J=7.2 Hz, 3H), 1.52 (s, 9H)

Preparation Example 40 Compound B-III

40-1) Preparation of Compound LX:

Tert-butyl 2-bromo-2-methylpropanoate (100 g, 0.6 mol), N-hydroxyphthalamide (136 g, 0.6 mol), and triethylamine (93 g, 0.9 mol) werestirred for 24 hours at 80° C. The resultant was diluted with ethylacetate (1 L×2), washed with water (1 L), 1N HCl (800 mL), and 0.5Nsodium hydroxide (500 mL), dehydrated with anhydrous sodium sulfate, andconcentrated under reduced pressure. The thus-obtained white solid waswashed with n-hexane (800 mL) and filtered under reduced pressure toyield Compound LX (91 g (49%)).

¹H NMR (600 MHz, chloroform-d₁) δ 7.85 (q, J=3 Hz, 2H), 7.76 (q, J=3 Hz,2H), 1.59 (s, 6H), 1.52 (s, 9H)

40-2) Preparation of Compound LXI:

Compound LX (2.63 g, 8.6 mmol) was dissolved in methylene chloride (11mL) and methanol (2 mL). Hydrazine monohydrate (1.7 mL) was added. Theresulting solution was stirred for 1 and a half hour at roomtemperature. The thus-obtained solid was filtered under reducedpressure. The filtrate was diluted with ethyl acetate (20 mL) and washedwith distilled water (20 mL×2) and saline (20 mL). The organic layer wasdehydrated with anhydrous sodium sulfate and concentrated under reducedpressure to yield Compound LXI (1.4 g, (93%)), which was used for nextstep without performing purification.

40-3) Preparation of Compound LXII:

Acetamidine hydrochloride (6 g, 64 mmol) was dissolved in distilledwater (75 mL) and the resulting solution was cooled to 0° C. Sodiumhyperchloride (4% chlorine available sol, 95 mL) was added over 1.5hours and the resulting solution was stirred for 1 hour. Excessiveamount of sodium chloride was added and the resultant was extracted withethyl acetate (150 mL×2). The organic layer was dehydrated withanhydrous sodium sulfate and concentrated under reduced pressure toyield Compound LXII (5.1 g (87%)), which was used for next step withoutperforming purification.

40-4) Preparation of Compound LXIII:

Compound LXII (5.1 g, 55 mmol) was dissolved in methanol (250 mL) andthe resulting solution was cooled to 0° C. Potassium thiocyanate (5.3 g,55 mmol) was then added. The resulting solution was stirred for 12 hoursat room temperature, concentrated under reduced pressure, diluted withethyl acetate (200 mL), and filtered under reduced pressure to remove asolid. The filtrate was concentrated under reduced pressure, creating asolid which was filtered under reduced pressure to yield Compound LXIII(2 g (32%)). The filtrate was further concentrated and applied to columnchromatography (SiO2, n-hex:EA=4:1) to yield Compound LXIII (2 g (32%)).

¹H NMR (600 MHz, CD₃OD) δ 3.27 (s, 3H)

40-5) Preparation of Compound LXIV:

Compound LXIII (2 g, 17.4 mmol) was added to Boc₂O (6 mL). The resultantwas stirred for 12 hours with reflux, concentrated under reducedpressure, and applied to column chromatography (SiO2, n-hex:EA=4:1) toyield Compound LXIV (2 g (53%)).

¹H NMR (600 MHz, chloroform-d₁) δ=10.97 (br, 1H), 2.55 (s, 3H), 1.55 (s,9H)

40-6) Preparation of Compound LXV:

Diisopropylamine (11.37 mL, 82 mmol) was dissolved in anhydroustetrahydrofuran (50 mL) and the resulting solution was cooled to −78.Butyl lithium (1.6 M n-hex sol., 56.1 mL, 90 mmol) was added and theresulting solution was stirred for 10 minutes at the same temperature toprepare LDA solution.

To the LDA solution, a solution of Compound LXIV (4.39 g, 20.4 mmol)dissolved in anhydrous tetrahydrofuran (20 mL) was slowly added at −78°C. Carbonic acid gas was introduced in the resulting solution −40° C.The resulting solution was stirred for 1 hour at −40° C., 30 minutes at0° C., and 4 hours at room temperature. Distilled water (5 mL) was addedto end the reaction. The resulting solution was concentrated underreduced pressure. By adding distilled water (200 mL) and extracting withether (150 mL×3), remaining starting material was recovered (2 g). Theresulting aqueous solution was acidified with 1N HCl aqueous solutionand extracted with ethyl acetate (250 mL). The organic layer wasdehydrated with anhydrous sodium sulfate and concentrated under reducedpressure to yield Compound LXV (2.2 g (40%)).

¹H NMR (600 MHz, DMSO-d6) δ 12.34 (s, 1H), 3.74 (s, 2H), 1.50 (s, 9H)

40-7) Preparation of Compound LXVI:

Compound LXV (2.2 g, 8.5 mmol) was dissolved in 1,4-dioxane (33 mL) andselenium dioxide (SeO2, 1.87 g, 17 mmol) was added. The resultingsolution was stirred at 100° C., cooled to room temperature, filteredunder reduced pressure, washed with 1,4-dioxane. The filtrate wasconcentrated under reduced pressure to yield Compound LXVI, which wasused for next step without performing purification.

40-8) Preparation of Compound LXVII:

Compound LXVII (12.9 g (64%)) was prepared by a method similar toPreparation Example 38-7 by using Compound LXVI (12.8 g, 47.1 mmol) andCompound LXI (10.2 g, 58.2 mmol).

¹H NMR (600 MHz, DMSO-d₆) δ 1.49 (s, 9H), 1.44 (s, 6H), 1.36 (s, 9H)

40-9) Preparation of Compound B-III:

Compound B-III (4.12 g (39%)) was prepared by a method similar toPreparation Example 38-8 by using Compound LXVII (5.85 g, 13.6 mmol) andCompound XLV (7.16 g, 17.6 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=8.75 (br, 1H), 8.05 (br, 1H), 7.35 (d,J=7.8 Hz, 2H), 6.96 (d, J=9 Hz, 2H), 6.09 (dd, J=4.8 Hz, 1H), 5.26 (d,J=11.4 Hz, 1H), 5.21 (d, J=11.4 Hz, 1H), 5.05 (d, J=4.8 Hz, 1H), 4.53(d, J=11.4 Hz, 1H), 4.47 (d, J=11.4 Hz, 1H), 3.82 (s, 3H), 3.65 (d, J=18Hz, 1H), 3.49 (d, J=18 Hz, 1H), 1.66 (s, 3H), 1.63 (s, 3H), 1.54 (s,9H), 1.40 (s, 9H)

Preparation Example 41 Compound LXXII

Compound LXXII was prepared by a method similar to Preparation Example38-7 by using Compound XLVII and Compound LXI.

Preparation Example 42 Compounds B-IV and B-V

42-1) Preparation of Compound LXVIII:

GCLE (49 g, 0.1 mol) was dissolved in acetone (1 L) and sodium iodide(45 g, 0.3 mol) was added. The resulting solution was stirred for 2hours at room temperature, concentrated under reduced pressure, dilutedwith ethyl acetate (1.2 L), and washed with water (500 ml), 10% sodiumthiosulfate (Na₂S₂O₃.5H₂O) aqueous solution (1 L), and saline (500mL×2). The organic layer was washed with anhydrous sodium sulfate andconcentrated under reduced pressure to yield Compound LXVIII (57 g(99%)), which was used for next step without performing purification.

42-2) Preparation of Compound LXIX:

Compound LXVIII (57 g, 0.1 mol) was dissolved in ethyl acetate (1 L) andtriphenylphosphine (52 g, 0.2 mol) was added at room temperature. Theresulting solution was stirred for 2 hours. The thus-created solid wasfiltered under reduced pressure and washed with ethyl acetate and driedto yield Compound LXIX (80 g (95%)).

¹H NMR (600 MHz, chloroform-d₁) δ=7.80˜7.64 (m, 15H), 7.35˜7.26 (m, 5H),7.16 (d, J=7.8 Hz, 2H), 7.65 (d, J=7.8 Hz, 2H), 6.46 (d, J=3.0 Hz, 1H),5.66 (dd, J=5.4 Hz, 9 Hz, 1H), 5.62 (t, J=15 Hz, 1H) 5.16 (t, J=15 Hz,1H), 4.58 (m, 3H), 4.05 (dd, 4.8 Hz, 18.6 Hz, 1H), 3.80 (s, 3H), 3.66(s, 2H), 3.37 (d, 18.6 Hz, 1H)

42-3) Preparation of Compound LXX:

Compound LXIX (40 g, 48 mmol) was dissolved in methylene chloride (450ml) and distilled water (150 mL). The resulting solution was cooled to0° C. and chloroacetaldehyde (50% aq sol, 30 mL, 238 mmol) was added.Then, 2N sodiumhydroxide aqueous solution (29 mL) was added and theresulting solution was stirred for 30 minutes at the same temperature.The organic layer was washed with water (200 mL) and saline (250 mL),dehydrated with anhydrous sodium sulfate, concentrated under reducedpressure, and applied to column chromatography (SiO2, n-hex:EA:MC=2:1:1)to yield Compound LXX (9.1 g (37%)).

¹H NMR (600 MHz, chloroform-d₁) δ=7.63 (m, 7H), 6.86 (d, J=9 Hz, 2H),6.20 (d, J=11.4 Hz, 1H), 6.01 (d, J=8.4 Hz, 1H), 5.82 (dd, 4.8 Hz, 9 Hz,1H), 5.72 (m, 1H), 5.14 (m, 2H), 4.98 (d, J=4.8 Hz, 1H), 3.91 (dd, J=8.4Hz, 12.6 Hz, 1H), 3.78 (s, 3H), 3.72 (dd, J=7.2 Hz, 12 Hz, 1H), 3.67 (q,J=16.2 Hz, 2H), 3.47 (dd, J=18.6 Hz, 124.8 Hz, 2H)

42-4) Preparation of Compound LXXI:

Compound LXXI (15 g (50%)) was prepared by a method similar toPreparation Example 36 by using Compound LXX (36 g, 70.2 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=7.31 (d, J=9.0 Hz, 2H), 6.87 (d, J=9Hz, 2H), 6.23 (d, J=11.4 Hz, 1H), 5.70 (m, 1H), 5.16 (m, 1H), 4.92 (d,J=4.8 Hz, 1H), 4.74 (d, J=4.8 Hz, 1H), 3.93 (dd, J=9.6 Hz, 11.4 Hz, 1H),3.78 (s, 3H), 3.72 (dd, J=7.2 Hz, 12 Hz, 1H), 3.51 (dd, J=18.6 Hz, 124.8Hz, 2H)

42-5) Preparation of Compound B-IV:

Compound B-IV (17 g (47%)) (E/Z mixture 2:8) was prepared by a methodsimilar to Preparation Example 38-8 by using Compound LXXII (19.7 g,34.5 mmol) and Compound LXXI (14.9 g, 34.5 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=8.18 (d, J=9 Hz, 1H), 7.33˜7.24 (m,17H), 6.89 (m, 3H), 6.71 (s, 1H), 6.27 (d, J=10.8 Hz, 1H), 5.98 (m, 1H),5.74 (m, 1H), 5.73 (m, 3H), 3.94 (dd, J=7.8 Hz, 9 Hz, 1H), 3.79 (s, 3H),3.75 (dd, J=7.8 Hz, 9 Hz, 1H), 3.47 (dd, J=18.Hz, 124.8 Hz, 2H), 1.62(d, J=27 Hz, 6H), 1.39 (s, 9H)

42-6) Preparation of Compound B-V:

Compound B-V (18 g (95%)) was prepared by a method similar toPreparation Example 42-1 by using Compound B-IV (17 g, 17.9 mmol) andused for next step without performing purification.

Preparation Example 43 Compounds B-VI and B-VII

43-1) Preparation of Compound B-VI:

Compound B-VI (2 g (52%)) (E/Z mixture 2:8) was prepared by a methodsimilar to Preparation Example 38-8 by using Compound LIV (2.5 g, 3.7mmol) and Compound LXXI (1.73 g, 3.7 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=8.16 (d, J=8.4 Hz, 1H), 7.35˜7.26 (m,27H), 6.98 (s, 1H), 6.91 (m, 3H), 6.75 (s, 1H), 6.33 (d, J=10.8 Hz, 1H),5.92 (dd, J=4.8 Hz, 8.4 Hz, 1H), 5.78 (m, 1H), 5.20 (m, 3H), 5.78 (d,J=4.8 Hz, 1H), 3.93 (dd, J=8.4 Hz, 12 Hz, 1H), 3.81 (s, 3H), 3.75 (dd,J=8.4 Hz, 12 Hz, 1H), 3.47 (dd, J=18 Hz, 89.4 Hz, 2H), 1.69 (d, J=7.8Hz, 3H)

43-2) Preparation of Compound B-VII:

Compound B-VII (1.1 g (99%)) was prepared by a method similar toPreparation Example 42-1 by using Compound B-VI (1 g, 0.96 mmol) andused for next step without performing purification.

Preparation Example 44 Compound B-VIII

Compound B-VIII (5.3 g (52%)) was prepared by a method similar toPreparation Example 38-8 by using Compound LXXII (6.8 g, 10 mmol) andCompound XLV (4.86 g, 12 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=8.23 (d, J=3.6 Hz, 1H), 7.33 (m, 17H),7.00 (s, 1H), 6.90 (m, 3H), 6.70 (s, 1H), 5.96 (dd, J=4.8 Hz, 8.4 Hz,1H), 5.24 (dd, J=11.4 Hz, 34.8 Hz, 2H), 5.00 (d, J=5.4 Hz, 1H), 5.51(dd, J=12 Hz, 50.4 Hz, 2H), 3.79 (s, 3H), 3.60 (d, J=18 Hz, 1H), 3.44(d, J=18 Hz, 1H), 1.61 (s, 3H), 1.59 (s, 3H), 1.39 (s, 9H)

Preparation Example 45 Compound B-IX

Compound B-VIII (5.0 g, 5.42 mmol) was dissolved in methylene chloride(50 mL) and m-chloroperbenzoic acid (0.84 g, 4.88 mmol) was added at−20° C. The resulting solution was stirred for 1 hour at 10° C. Sodiumthiosulfate saturated aqueous solution (30 mL) was then added. Theresulting solution was ⅓ concentrated under reduced pressure, extractedwith ethyl acetate (100 mL×2), dehydrated with anhydrous sodium sulfate,concentrated under reduced pressure, and applied to columnchromatography (EA:Hex=1:3-1:2) to yield Compound B-IX (3.73 g (73%)).

¹H NMR (600 MHz, CDCl₃) δ 7.87 (d, J=9.6 Hz, 1H), 7.36 (d, J=8.4 Hz,2H), 7.31˜7.27 (m, 15H), 7.03 (s, 1H), 6.92 (d, J=9 Hz, d), 6.69 (s,1H), 6.21 (q, J=4.8 Hz, 1H), 5.29 (d, J=11.4 Hz, 1H), 5.24 (d, J=11.4H),5.07 (d, J=12 Hz, 1H), 4.55 (d, J=4.8 Hz), 4.22 (d, J=12.6 Hz, 1H), 3.82(s, 3H), 3.74 (d, J=19.2 Hz, 1H), 3.39 (d, J=18.6 Hz, 1H), 1.58 (d, J=15Hz, 6H), 1.41 (s, 9H)

Preparation Example 46 Compound B-X

46-1) Preparation of Compound LXXIII:

Compound LXXIII was prepared by a method similar to Preparation Example38-7 by using Compound LVIII and Compound LXI.

46-2) Preparation of Compound B-X:

Compound B-X (4 g (25%)) was prepared by a method similar to PreparationExample 38-8 by using Compound LXXIII (9.2 g, 19.8 mmol) and CompoundXLV (10.45 g, 25.8 mmol).

¹H NMR (600 MHz, chloroform-d₁) δ=7.92 (d, J=9.0 Hz, 1H), 7.87 (brs,1H), 7.36 (d, J=9.6 Hz, 2H), 6.92 (d, J=9.6 Hz, 2H), 6.05 (dd, J=5.4 Hz,9.6 Hz, 1H), 5.28 (dd, J=11.4 Hz, 36.6 Hz, 2H), 5.04 (d, J=5.4 Hz, 1H),4.56 (dd, J=12 Hz, 56.4 Hz, 2H), 3.82 (s, 3H), 3.66 (dd, J=18 Hz, 97.8Hz, 2H), 1.62 (s, 3H), 1.60 (s, 3H), 1.42 (s, 9H)

Preparation Example 47 Compound B-XI

Compound B-XI was prepared by a method similar to Preparation Example 43by using Compound LXVII and Compound LXXI.

¹H NMR (600 MHz, chloroform-d₁) δ=8.98 (brs, 1H), 8.06 (d, J=9.0 Hz,1H), 7.31 (d, J=8.4 Hz, 2H), 6.87 (d, J=8.4 Hz, 1H), 6.27 (d, J=11.4 Hz,1H), 6.07 (m, 1H), 5.74 (m, 1H), 5.14 (m, 3H), 3.93˜3.71 (m, 2H), 3.52(m, 2H), 1.62 (s, 3H), 1.60 (s, 3H), 1.52 (s, 9H): NMR of allylchloride.

Preparation Example 48 Compound B-XII

Compound B-XII was prepared by a method similar to Preparation Example43 by using Compound LIX and Compound LXXI.

¹H NMR (600 MHz, chloroform-d₁) δ=7.99 (brs, 1H), 7.85 (d, J=9.0 Hz,1H), 7.36˜7.12 (m, 12H), 6.92 (d, J=8.4 Hz, 1H), 6.31 (d, J=11.4 Hz,1H), 6.04 (m, 1H), 5.78 (m, 1H), 5.15˜4.99 (m, 3H), 3.94˜3.72 (m, 2H),3.45 (m, 2H), 1.66 (t, J=3 Hz, 3H), 1.52 (s, 9H): NMR of allylchloride.

Preparation Example 49 Compound B-XIII

Compound B-XIII was prepared by a method similar to Preparation Example43-3 by using Compound LXXIII and Compound LXXI.

¹H NMR (600 MHz, chloroform-d₁) δ=7.91 (m, 2H), 7.36 (d, J=8.4 Hz, 2H),6.91 (d, J=8.4 Hz, 1H), 6.30 (d, J=11.4 Hz, 1H), 6.06 (m, 1H), 5.77 (m,1H), 5.20 (dd, J=12 Hz, 22.8 Hz, 2H), 5.10 (d, J=4.8 Hz, 1H), 3.96˜3.74(m, 2H), 3.55 (dd, J=18 Hz, 99.6 Hz, 2H), 1.62 (s, 3H), 1.60 (s, 3H),1.42 (s, 9H): NMR of allylchloride.

Example 1 Compound 1

Compound B-VIII (193 mg, 0.21 mmol) was dissolved inN,N-dimethylformamide (0.5 mL) and sodium iodide (31 mg, 0.21 mmol) wasadded. The resulting solution was stirred for 30 minutes at roomtemperature. Compound A-II (86 mg, 0.21 mmol) was added. Then, theresulting solution was stirred for 4 hours at room temperature, dilutedwith ethyl acetate (5 mL), washed with water (3 mL) and saline (3 mL),dehydrated with anhydrous sodium sulfate, concentrated under reducedpressure, and applied to column chromatography (MC:MeOH=50:1˜10:1) toyield a quaternary salt compound (67 mg (22%)).

The quaternary salt compound (67 mg, 40 umol) was dissolved in methylenechloride (0.5 mL). Anisole (0.2 mL) and trifluoroacetic acid (0.5 mL)were sequentially added. The resulting solution was stirred for 4 hoursat room temperature. Isopropyl ether (5 mL) was added. The thus-createdsolid was filtered under reduced pressure to yield Compound 1 (34 mg(94%)).

¹H NMR (600 MHz, DMSO-d₆+D2O) δ 8.36 (s, 1H), 7.86 (s, 1H), 7.61 (s,1H), 7.58 (s, 1H), 6.77 (s, 1H), 5.90 (d, J=4.8 Hz, 1H), 5.21 (d, J=4.8Hz, 1H), 5.09 (dd, J=15 Hz, 33.6 Hz, 2H), 3.60 (m, 4H), 3.28 (m, 2H),1.43 (s, 3H), 1.42 (s, 3H)

Example 2 Compound 2

Compound 2 (2.6 mg, 8%) was prepared by a method similar to Example 1 byusing Compound B-VIII and Compound A-II.

¹H NMR (600 MHz, CD₃OD) δ 8.40 (s, 1H), 7.90 (s, 1H), 7.83 (s, 1H), 7.34(s, 1H), 7.03 (s, 1H), 5.95 (d, J=4.8 Hz, 1H), 5.29 (m, 3H), 3.93 (s,3H), 3.66˜3.37 (m, 6H), 1.61 (s, 3H), 1.60 (s, 3H)

Example 3 Compound 3

Compound 3 (25 mg, 41%) was prepared by a method similar to Example 1 byusing Compound B-I and Compound A-II.

¹H NMR (600 MHz, DMSO-d₆+D2O) δ 8.33 (s, 1H), 7.79 (s, 1H), 7.59 (s,1H), 7.53 (s, 1H), 6.72 (s, 1H), 5.87 (d, J=4.8 Hz, 1H), 5.14 (d, J=4.8Hz, 1H), 5.05 (m, 2H), 4.54 (q, J=7.2 Hz, 1H), 3.54˜3.22 (m, 6H), 1.35(s, 3H), 1.32 (s, 3H)

Example 4 Compound 4

Compound 4 (10 mg, 28%) was prepared by a method similar to Example 1 byusing Compound B-II and Compound A-II.

¹H NMR (600 MHz, DMSO-d₆+D2O) δ 8.41 (s, 1H), 7.85 (s, 1H), 7.71 (s,1H), 7.60 (s, 1H), 5.87 (d, J=5.4 Hz, 1H), 5.16 (d, J=4.8 Hz, 1H), 5.08(m, 2H), 4.58 (q, J=7.2 Hz, 1H), 3.59˜3.26 (m, 6H), 1.41 (d, J=7.2 Hz,3H)

Example 5 Compound 5

Compound 5 (7.6 mg, 24%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-IV.

¹H NMR (600 MHz, CD₃OD) δ 8.52 (s, 1H), 7.88 (s, 1H), 7.86 (s, 1H), 7.61(s, 1H), 7.06 (s, 1H), 5.97 (d, J=5.4 Hz, 1H), 5.32 (d, J=14.4 Hz, 1H),5.26 (d, J=5.4 Hz, 1H), 5.17 (d, J=14.4 Hz, 1H), 3.98 (m, 2H), 3.70 (m,2H), 3.44 (m, 2H), 3.36˜3.28 (m, 4H), 1.58 (s, 3H), 1.56 (s, 3H)

Example 6 Compound 6

Compound 6 (12 mg, 17%) was prepared by a method similar to Example 1 byusing Compound B-VIII and Compound A-V.

¹H NMR (600 MHz, DMSO-d⁶) δ 11.72 (s, 1H), 11.05 (s, 1H), 10.88 (s, 1H),9.48 (d, J=8.4 Hz, 1H), 8.67 (br, 1H), 8.48 (s, 1H), 7.79 (s, 1H), 7.55(s, 1H), 7.31 (br, 2H), 6.70 (s, 1H), 5.98 (br, 1H), 5.93 (dd, J=4.8 Hz,5.4 Hz, 1H), 5.21 (d, J=4.8 Hz, 1H), 5.12 (d, J=14.4 Hz, 1H), 4.94 (d,J=15 Hz, 1H), 3.75 (m, 2H), 3.60 (m, 2H), 3.42 (m, 2H), 1.41 (s, 3H),1.40 (s, 3H)

Example 7 Compound 7

Compound 7 (38 mg, 39%) was prepared by a method similar to Example 1 byusing Compound B-VIII and Compound A-VI.

¹H NMR (600 MHz, DMSO-d⁶+D₂O) δ 8.36 (s, 1H), 7.84 (s, 1H), 7.57 (s,1H), 7.51 (s, 1H), 6.73 (s, 1H), 5.91 (d, J=4.8 Hz, 1H), 5.20 (d, J=4.8Hz, 1H), 5.08 (q, J=15.6 Hz, 2H), 3.54˜3.41 (m, 2H), 3.37 (br, 2H), 3.05(br, 2H), 1.64 (br, 4H), 1.43 (s, 3H), 1.41 (s, 3H)

Example 8 Compound 8

Compound B-V (371 mg, 0.39 mmol) and Compound A-II (200 mg, 0.36 mmol)were dissolved in N,N-dimethylformamide (2 mL). The resulting solutionwas stirred for 3 hours at room temperature, diluted with ethyl acetate(15 mL), washed with water (20 mL) and saline (10 mL), dehydrated withanhydrous sodium sulfate, concentrated under reduced pressure, andapplied to column chromatography (MC:MeOH=50:1˜10:1) to yield aquaternary salt compound (323 mg (60%)).

The quaternary salt compound (323 mg, 0.22 mmol) was dissolved inmethylene chloride (4 mL). Anisole (0.5 mL) and trifluoroacetic acid (4mL) were sequentially added. The resulting solution was stirred for 3hours at room temperature. Isopropyl ether (20 mL) was added and thethus-created solid was filtered under reduced pressure to yield Compound8 (176 mg, 100%).

¹H NMR (600 MHz, DMSO-d⁶+D₂O) δ 8.40 (s, 1H), 7.85 (s, 1H), 7.67 (s,1H), 7.58 (s, 1H), 6.91 (d, J=16.2 Hz, 1H), 6.77 (s, 1H), 6.31 (m, 1H),5.84 (d, J=4.8 Hz, 1H), 5.22 (d, J=4.8 Hz, 1H), 4.92 (m, 2H), 3.85 (d,J=18 Hz, 1H), 3.74 (d, J=18 Hz, 1H), 3.57 (m, 2H), 3.30 (br, 2H), 1.45(s, 3H), 1.43 (s, 3H)

Example 9 Compound 9

Compound 9 (35 mg, 28%) was prepared by a method similar to Example 8 byusing Compound B-VII and Compound A-II.

¹H NMR (600 MHz, DMSO-d6) δ 11.78 (t, J=6.6 Hz, 1H), 11.03 (br d, 2H),9.54 (d, J=8.4 Hz, 1H), 9.00 (s, 1H), 8.42 (s, 1H), 8.09 (br, 1H), 7.82(s, 1H), 7.70 (s, 1H) 7.57 (s, 1H) 7.34 (br, 2H) 6.93 (d, J=15.6 Hz),6.78 (s, 1H), 6.31 (m, 1H), 6.04 (br, 1H), 5.87 (dd, J=4.8 Hz, 8.4 Hz,1H), 5.22 (d, J=4.8 Hz, 1H), 4.93 (m, 1H), 4.62 (q, J=6.6 Hz, 1H),3.83˜3.29 (m, 6H), 1.4 (d, J=7.2 Hz, 3H)

Example 10 Compound 10

Compound 10 (8 mg, 3.2%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-II.

¹H NMR (600 MHz, CD₃OD) δ 8.34 (s, 1H), 7.93 (s, 1H), 7.58 (s, 1H), 7.10(s, 1H), 7.02 (s, 1H), 6.97 (d, J=16.2 Hz, 1H), 6.23 (m, 1H), 5.92 (d,J=4.8 Hz, 1H), 5.21 (d, J=4.8 Hz, 1H), 4.96 (m, 2H), 4.42 (s, 2H), 3.83(d, J=18 Hz, 1H), 3.66˜3.61 (m, 3H), 3.41 (t, J=5.4 Hz), 1.62 (s, 3H),1.59 (s, 3H)

Example 11 Compound 11

Compound 11 (51 mg, 30%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-VIII.

¹H NMR (600 MHz, DMSO-d6) δ 9.44 (d, J=9 Hz, 1H), 8.97 (br, 1H), 8.67(br, 1H), 8.39 (s, 1H), 8.01 (br, 1H), 7.90 (s, 1H), 7.63 (s, 1H), 7.41(s, 1H), 7.34 (br, 2H), 6.89 (d, J=15.6 Hz, 1H), 6.69 (s, 1H), 6.27 (m,1H), 5.93 (br, 1H), 5.81 (dd, J=4.8 Hz, 8.4 Hz, 1H), 5.18 (d, J=4.8 Hz,1H), 4.89 (m, 2H), 3.81 (d, J=17.4 Hz, 1H), 3.55˜3.21 (m, 5H), 1.40 (s,3H), 1.39 (s, 3H)

Example 12 Compound 12

Compound 12 (40 mg, 39%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-X.

¹H NMR (600 MHz, CD₃OD) δ 8.66 (s, 1H), 8.60 (s, 1H), 7.90 (s, 1H), 7.65(s, 1H), 7.19 (d, J=15.6 Hz, 1H), 6.99 (s, 1H), 6.22 (m, 1H), 5.92 (d,J=4.8 Hz, 1H), 5.23 (d, J=4.8 Hz, 1H), 4.93 (m, 2H), 3.82˜3.77 (m, 3H),3.68 (d, J=18 Hz, 1H), 2.80 (t, J=6 Hz, 2H), 1.61 (s, 3H), 1.60 (s, 3H)

Example 13 Compound 13

Compound 13 (70 mg, 69%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-XI.

¹H NMR (600 MHz, CD₃OD) δ 8.62 (s, 1H), 8.60 (s, 1H), 7.90 (s, 1H), 7.45(s, 1H), 7.21 (d, J=16.2 Hz, 1H), 7.06 (s, 1H), 6.22 (m, 1H), 5.94 (d,J=4.8 Hz, 1H), 5.23 (d, J=4.8 Hz, 1H), 4.90 (m, 2H) 3.83˜3.65 (m, 4H),2.80 (m, 2H), 1.63 (s, 3H), 1.61 (s, 3H)

Example 14 Compound 14

Compound 14 (88 mg, 42%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-XII.

¹H NMR (600 MHz, CD₃OD) δ 8.66 (s, 1H), 8.65 (s, 1H), 7.97 (s, 1H), 7.69(s, 1H), 7.23 (d, J=16.2 Hz, 1H), 7.03 (s, 1H), 6.25 (m, 1H), 5.94 (d,J=4.2 Hz, 1H), 5.23 (d, J=4.8 Hz, 1H), 4.92 (m, 2H), 3.85 (d, J=18 Hz,1H), 3.69 (d, J=17.4 Hz), 1.62 (s, 3H), 1.61 (s, 3H)

Example 15 Compound 15

Compound 15 (65 mg, 12%) was prepared by a method similar to Example 1by using Compound B-X and Compound A-II.

¹H NMR (600 MHz, CD₃OD) δ 8.37 (s, 1H), 8.28 (s, 1H), 7.87 (s, 1H), 7.65(s, 1H), 5.95 (d, J=4.8 Hz, 1H), 5.28 (d, J=14.4 Hz, 1H), 5.24 (d, J=5.4Hz, 1H), 5.10 (d, J=15 Hz, 1H), 3.69 (m, 3H), 3.43˜3.33 (m, 3H), 1.56(s, 3H), 1.55 (s, 3H)

Example 16 Compound 16

Compound 16 (25 mg, 9%) was prepared by a method similar to Example 1 byusing Compound B-III and Compound A-II.

¹H NMR (600 MHz, DMSO-d₆+D₂O) δ 8.31 (s, 1H), 7.80 (s, 1H), 7.79 (s,1H), 7.52 (s, 1H), 5.84 (d, J=4.8 Hz, 1H), 5.12 (d, J=4.8 Hz, 1H), 5.05(m, 2H), 3.55˜3.20 (m, 6H), 1.37 (s, 6H)

Example 17 Compound 17

Compound 17 (30 mg, 16%) was prepared by a method similar to Example 1by using Compound B-II and Compound A-VIII.

¹H NMR (600 MHz, CD₃OD) δ 8.39 (d, J=1.8 Hz, 1H), 8.00 (s, 1H), 7.89 (d,J=1.8 Hz, 1H), 7.60 (s, 1H), 5.96 (d, J=4.8 Hz, 1H), 5.34 (d, J=14.4 Hz,1H), 5.24 (d, J=4.8 Hz, 1H), 4.97 (d, J=15 Hz, 1H), 4.81 (q, J=7.2 Hz,1H), 3.65˜3.33 (m, 6H), 1.51 (d, J=7.8 Hz, 3H)

Example 18 Compound 18

Compound 18 (69 mg, 20%) was prepared by a method similar to Example 1by using Compound B-X and Compound A-X.

¹H NMR (600 MHz, CD₃OD) δ 8.78 (s, 1H), 8.67 (s, 1H), 7.91 (s, 1H), 7.65(s, 1H), 5.95 (d, J=4.8 Hz, 1H), 5.31 (d, J=15 Hz, 1H), 5.24 (d, J=4.8Hz), 5.14 (d, J=15.6 Hz, 1H), 3.80 (m, 2H), 3.70 (d, J=18 Hz, 1H), 3.14(d, J=18 Hz, 1H), 2.78 (m, 2H), 1.59 (s, 3H), 1.57 (s, 3H)

Example 19 Compound 19

Compound 19 (134 mg, 48%) was prepared by a method similar to Example 1by using Compound B-III and Compound A-X.

¹H NMR (600 MHz, DMSO-d₆+D2O) δ 8.77 (s, 1H), 8.64 (s, 1H), 7.82 (s,1H), 7.56 (s, 1H), 5.91 (d, J=4.8 Hz, 1H), 5.20˜4.97 (m, 3H), 3.56 (m,4H), 2.69 (m, 2H), 1.43 (s, 6H)

Example 20 Compound 20

Compound 20 (120 mg, 20%) was prepared by a method similar to Example 1by using Compound B-II and Compound A-X.

¹H NMR (600 MHz, DMSO-d₆+D2O) δ 8.76 (s, 1H), 8.66 (s, 1H), 7.82 (s,1H), 7.53 (s, 1H), 5.65 (d, J=4.2 Hz, 1H), 5.09 (d, J=14.4 Hz, 1H), 4.92(d, J=4.8 Hz, 1H), 4.86 (d, J=4.8 Hz, 1H), 4.58 (q, J=14.4 Hz, 1H), 3.56(m, 2H), 3.41 (d, J=18 Hz, 1H), 3.20 (d, J=17.4 Hz, 1H), 1.36 (s, 3H),1.35 (s, 3H)

Example 21 Compound 21

Compound 21 (145 mg, 53%) was prepared by a method similar to Example 1by using Compound B-II and Compound A-IV.

¹H NMR (600 MHz, CD₃OD) δ 8.45 (s, 1H), 7.93 (s, 1H), 7.86 (s, 1H), 7.61(s, 1H), 5.98 (d, J=4.8 Hz, 1H), 5.33 (d, J=14.4 Hz, 1H), 5.24 (d, J=4.8Hz, 1H), 5.18 (d, J=15 Hz, 1H), 4.81 (q, J=7.2 Hz, 1H), 4.12˜3.33 (m,10H), 1.53 (d, J=3.6 Hz, 3H)

Example 22 Compound 22

Compound 22 (33 mg, 8%) was prepared by a method similar to Example 1 byusing Compound B-X and Compound A-XII.

¹H NMR (600 MHz, CD₃OD) δ 8.81 (s, 1H), 8.75 (s, 1H), 7.97 (s, 1H), 7.70(s, 1H), 5.97 (d, J=4.8 Hz), 5.35 (d, J=14.4 Hz, 1H), 5.24 (d, J=5.4 Hz,1H), 5.01 (m, 1H), 3.49˜3.43 (m, 2H), 1.60 (br, 6H)

Example 23 Compound 23

Compound 23 (11 mg, 28%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-VI.

¹H NMR (600 MHz, DMSO-d6) δ 11.72 (t, J=6.6 Hz, 1H), 11.03 (br, 1H),10.83 (br, 1H), 9.48 (d, J=8.4 Hz, 1H), 8.98 (s, 1H), 8.58 (s, 1H), 8.15(br, 1H), 7.81 (s, 1H), 7.56 (s, 1H) 7.51 (s, 1H) 7.33 (br, 2H) 6.85 (d,J=15.6 Hz, 1H), 6.74 (s, 1H), 6.28 (m, 1H), 5.87 (dd, J=5.4 Hz, 8.4 Hz,1H), 5.79 (br, 1H), 5.23 (d, J=4.8 Hz, 1H), 4.93 (m, 2H), 3.84 (d,J=17.4 Hz, 1H), 3.62 (d, J=7.8 Hz 1H), 3.5˜3.09 (m, 4H), 1.66 (m, 4H),1.44 (s, 3H) 1.43 (s, 3H)

Example 24 Compound 24

Compound 24 (49 mg, 38%) was prepared by a method similar to Example 8by using Compound B-XII and Compound A-II.

¹H NMR (600 MHz, DMSO-d⁶) δ 11.76 (t, J=5.4 Hz, 1), 11.05 (s, 1H), 10.90(s, 1H), 9.53 (d, J=8.4 Hz, 1H), 9.00 (s, 1H), 8.42 (s, 1H), 8.09 (s,1H), 7.82 (s, 1H), 7.69 (s, 1H), 7.57 (s, 1H), 7.42 (br, 2H), 7.12 (t,J=7.2 Hz, 1H), 6.91 (d, J=16.2 Hz, 1H), 6.30 (m, 1H), 6.05 (br, 1H),5.84 (dd, J=4.8 Hz, 5.4 Hz, 1H), 5.20 (d, J=4.8 Hz, 1H), 4.91 (m, 2H),4.63 (q, J=7.2 Hz, 1H), 3.82 (d, J=18 Hz, 1H), 3.58 (m, 3H), 3.30 (m,2H), 1.42 (d, J=6.6 Hz, 3H)

Example 25 Compound 25

Compound 25 (26 mg, 36%) was prepared by a method similar to Example 8by using Compound B-XI and Compound A-II.

¹H NMR (600 MHz, DMSO d-₆) δ 11.74 (t, J=5.4 Hz, 1H), 9.52 (d, J=7.8 Hz,1H), 8.97 (bs, 1H), 8.44 (s, 1H), 8.20 (bs, 2H), 8.06 (bs, 2H), 7.81 (s,1H), 7.63 (s, 1H), 7.11 (m, 1H), 6.15 (bs, 2H), 5.75 (m, 1H), 5.07 (m,1H), 4.94 (m, 1H), 4.80 (m, 1H), 3.73 (m, 2H), 3.55 (m, 4H), 1.46 (s,3H), 1.45 (s, 3H)

Example 26 Compound 26

Compound 26 (22 mg, 29%) was prepared by a method similar to Example 8by using Compound B-XIII and Compound A-II.

¹H NMR (600 MHz, DMSO d-₆) δ 11.77 (t, J=5.4 Hz, 1H), 11.04 (bs, 1H),10.91 (bs, 1H), 9.45 (d, J=8.4 Hz, 1H), 9.00 (bs, 1H), 8.42 (s, 1H),8.09 (bs, 1H), 7.82 (s, 1H), 7.70 (s, 1H), 7.57 (s, 1H), 7.42 (s, 1H),6.90 (d, J=16.2 Hz, 1H), 6.29 (m, 1H), 6.05 (t, J=4.8 Hz, 1H), 5.82 (dd,J₁=8.4 Hz, J₂=5.4 Hz, 1H), 5.20 (d, J=5.4 Hz, 1H), 4.88 (m, 2H), 3.83(d, J=18.0 Hz, 1H), 3.58 (m, 3H), 3.29 (m, 2H), 1.47 (s, 3H), 1.45 (s,3H)

Example 27 Compound 27

Compound 27 (6 mg, 8%) was prepared by a method similar to Example 1 byusing Compound B-VIII and Compound A-XIII.

¹H NMR (600 MHz, CD₃OD) δ=8.37 (br, 1H), 7.91 (br, 1H), 7.34 (br, 1H),6.79 (br, 1H), 6.60 (br, 1H), 5.95 (d, 1.8 Hz, 1H), 5.25 (m, 2H), 4.94(br, 1H), 3.73˜3.23 (m, 6H), 3.10 (d, J=6.6 Hz, 3H), 1.66 (d, J=3.6 Hz,6H)

Example 28 Compound 28

Compound 28 (17 mg, 33%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-XIV.

¹H NMR (600 MHz, CD₃OD) δ 8.76 (s, 1H), 8.66 (s, 1H), 7.25 (d, J=2.4 Hz,1H), 7.18 (dd, J=2.4 Hz, 8.4 Hz, 1H), 7.04 (s, 1H), 6.77 (d, J=8.4 Hz,1H), 5.95 (d, J=5.4 Hz, 1H) 5.29 (d, J=15.6 Hz, 1H), 5.21 (d, J=4.8 Hz,1H), 4.94 (d, J=15.6 Hz, 1H), 3.69 (m, 3H), 3.41 (d, J=18.6 Hz, 1H),2.72 (t, J=6.6 Hz, 2H), 1.61 (s, 6H)

Example 29 Compound 29

Compound 29 (42 mg30%) was prepared by a method similar to Example 1 byusing Compound B-VIII and Compound A-XV.

¹H NMR (600 MHz, DMSO-d₆) δ=9.62 (br, 1H), 9.48 (d, J=6.6 Hz, 1H), 9.23(s, 1H), 8.43 (s, 1H), 8.23 (br, 1H), 7.61 (s, 1H), 7.37 (br, 1H), 6.87(s, 1H), 6.79˜6.74 (m, 3H), 6.70 (s, 1H), 6.17 (br, 1H), 5.91 (m, 1H),5.18 (d, J=4.8 Hz, 1H), 5.06 (br, 2H), 4.25 (br, 2H), 4.10 (br, 2H),3.48 (br, 2H), 3.34 (br, 1H), 3.23 (br, 1H), 2.71 (s, 3H), 1.42 (d,J=6.6 Hz, 6H)

Example 30 Compound 30

Compound 30 (12 mg, 14%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-XVI.

¹H NMR (600 MHz, DMSO-d₆) δ 9.51 (br, 1H), 9.46 (d, J=8.4 Hz, 1H), 8.35(s, 1H), 8.25 (t, J=4.2 Hz, 1H), 8.13 (s, 1H), 7.61 (s, 1H), 7.35 (br,2H), 7.26 (d, J=6.6 Hz, 1H), 7.15 (dd, J=2.4 Hz, 7.8 Hz, 1H), 6.73 (d,J=8.4 Hz, 1H), 6.675 (s, 1H), 5.94 (br, 1H), 5.86 (dd, J=8.4 Hz, 5.4 Hz,1H), 5.15 (d, J=5.4 Hz, 5.07 (m, 2H), 3.5˜3.34 (m, 4H), 3.18 (m, 2H),1.38 (s, 3H), 1.36 (s, 3H)

Example 31 Compound 31

Compound 31 (10 mg, 16%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-XVIII.

¹H NMR (600 MHz, DMSO-d₆) δ 9.49 (d, J=8.4 Hz, 1H), 9.09 (br, 1H) 8.38(s, 1H), 8.25 (br, s) 8.25 (t, J=5.4 Hz, 1H), 7.54 (s, 1H), 7.34 (br,2H), 7.27 (d, J=1.8 Hz, 1H), 7.15 (dd, J=1.8 Hz, 6.6 Hz, 1H), 6.76 (d,J=7.8 Hz, 1H), 6.71 (s, 1H), 5.92 (dd, J=4.8 Hz, 8.4 Hz, 1H), 5.84 (br,1H), 5.19 (d, J=4.8 Hz, 5.05 (m, 2H), 3.5˜3.32 (m, 4H), 3.07 (m, 2H),1.88 (m, 2H), 1.42 (s, 3H), 1.41 (s, 3H)

Example 32 Compound 32

Compound 32 (65 mg, 24%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-XIX.

¹H NMR (600 MHz, DMSO-d₆+D2O) δ 8.76 (s, 1H), 8.73 (s, 1H), 7.30 (d,J=2.4 Hz, 1H), 7.25 (dd, J=2.4 Hz, 8.4 Hz, 1H), 6.83 (dd, J=1.2 Hz, 7.8Hz, 6.76 (s, 1H), 5.90 (4.2 Hz, 1H), 5.20 (d, J=4.8 Hz, 1H), 5.14 (d,J=15 Hz, 1H), 4.99 (d, J=15 Hz, 1H), 4.08 (s, 2H), 3.60 (dd, J=18 Hz,59.4 Hz, 2H), 1.46 (s, 3H), 1.44 (s, 3H)

Example 33 Compound 33

Compound 33 (15 mg, 12%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-XX.

¹H NMR (600 MHz, DMSO-d₆, D₂O) δ 8.34 (s, 1H), 8.21 (br, s, 1H) 7.77 (d,J=6 Hz, 1H), 7.73 (s, 1H), 6.81 (d, J=8.4 Hz, 1H), 6.67 (s, 1H), 5.77(d, J=4.2 Hz, 1H), 5.10 (d, J=4.2 Hz, 1H), 5.04˜4.86 (m, 2H), 3.55˜3.20(m, 6H), 1.39 (s, 3H), 1.37 (s, 3H)

Example 34 Compound 34

Compound 34 (10 mg, 9%) was prepared by a method similar to Example 1 byusing Compound B-I and Compound A-XXI.

¹H NMR (600 MHz, CD3OD) δ 8.33 (s, 1H), 7.83 (s, 1H), 7.25 (s, 1H), 7.20(d, J=7.8 Hz, 1H), 6.97 (s, 1H), 6.79 (d, J=8.4 Hz, 1H), 5.92 (d, J=4.8Hz, 1H), 5.21˜(m, 2H), 4.92 (m, 2H), 3.72˜3.67 (m, 6H), 1.51 (2, J=7.2Hz, 3H)

Example 35 Compound 35

Compound 35 (49 mg, 31%) was prepared by a method similar to Example 1by using Compound B-II and Compound A-XXI.

¹H NMR (600 MHz, DMSO-d₆) δ 9.54 (m, 2H) 9.14 (s, 1H), 9.06 (s, 1H),8.36 (s, 1H), 8.30 (t, J=5.4 Hz, 1H), 8.15 (s, 1H), 7.75 (s, 1H), 7.42(s, 1H), 7.32 (s, 1H), 7.19 (dd, J=1.8 Hz, 7.8 Hz, 1H), 6.76 (d, J=7.8Hz, 1H), 5.98 (t, J=5.4 Hz, 1H), 5.87 (dd, J=5.4 Hz, 7.8 Hz, 1H), 5.15(d, J=4.8 Hz, 1H), 5.08 (m, 2H), 4.60 (q, J=7.2 Hz, 1H), 3.52˜3.20 (m,6H), 1.40 (d, J=6.6 Hz, 3H)

Example 36 Compound 36

Compound 36 (12 mg, 52%) was prepared by a method similar to Example 1by using Compound B-VIII and 4,5-diaminopyrimidine.

¹H NMR (600 MHz, CD₃OD) δ 8.34 (d, J=1.8 Hz, 1H), 7.78 (d, J=1.8 Hz,1H), 7.02 (s, 1H), 5.99 (d, J=4.8 Hz, 1H) 5.30˜4.90 (m, 3H), 3.65 (d,J=18 Hz, 1H), 3.34 (d, J=18 Hz, 1H), 1.60 (s, 6H)

Example 37 Compound 37

Compound 37 (19 mg, 31%) was prepared by a method similar to Example 1by using Compound B-II and Compound A-I.

¹H NMR (600 MHz, CD₃OD) δ 8.43 (s, 1H), 7.94 (s, 1H), 7.04 (s, 1H), 5.90(d, J=4.8 Hz, 1H), 5.25˜4.8 (m, 3H), 3.70 (d, J=18.6 Hz, 1H), 3.49 (t,J=6 Hz, 1H), 3.30˜3.25 (m, 3H), 1.62 (s, 6H)

Example 38 Compound 38

Compound 38 (7 mg, 9%) was prepared by a method similar to Example 1 byusing Compound B-VIII and Compound A-XXII.

¹H NMR (600 MHz, CD₃OD) δ=8.36 (s, 1H), 7.70 (s, 1H), 7.04 (s, 1H), 5.98(d, J=4.8 Hz, 1H), 5.27˜4.8 (m, 3H), 3.70˜3.50 (m, 2H), 3.49 (m, 2H),2.45 (m, 2H), 1.98 (m, 2H), 1.62 (s, 6H)

Example 39 Compound 39

Compound 39 (2 mg, 2%) was prepared by a method similar to Example 1 byusing Compound B-VIII and Compound A-XXIII.

¹H NMR (600 MHz, CD₃OD) δ 8.42 (d, J=1.8 Hz, 1H), 7.70 (d, J=1.8 Hz,1H), 6.99 (s, 1H), 5.92 (d, J=4.8 Hz, 1H) 5.27˜4.90 (m, 3H), 3.63 (d,J=18 Hz, 1H), 3.34 (d, J=18 Hz, 1H), 3.15 (d, J=3.6 Hz, 3H), 1.62 (s,6H)

Example 40 Compound 40

Compound 40 (12 mg, 18%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-XXIV.

¹H NMR (600 MHz, CD₃OD) δ 8.41 (d, J=1.8 Hz, 1H), 7.98 (d, J=1.8 Hz,1H), 7.03 (s, 1H), 5.91 (d, J=4.8 Hz, 1H), 5.23˜4.90 (m, 3H), 3.68˜3.37(m, 6H), 1.60 (s, 6H)

Example 41 Compound 41

Compound 41 (70 mg, 40%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-XXVI.

¹H NMR (600 MHz, DMSO-d⁶+D₂O) δ 8.46 (s, 1H), 7.49 (s, 1H), 6.74 (s,1H), 5.90 (d, J=4.2 Hz, 1H), 5.20 (d, J=4.8 Hz, 1H), 5.10 (q, J=15.6 Hz,2H), 3.62˜3.40 (m, 6H), 3.03 (br, 2H), 2.83 (br, 4H), 1.66˜1.59 (m, 8H),1.44 (s, 3H), 1.43 (s, 3H)

Example 42 Compound 42

Compound 42 (6.2 mg, 28%) was prepared by a method similar to Example 8by using Compound B-XII and Compound A-XXVII.

¹H NMR (600 MHz, CD₃OD) δ 8.26 (s, 1H), 7.41 (s, 1H), 7.09 (d, J=16.2Hz, 1H), 6.25 (m, 1H), 5.91 (d, J=4.8 Hz, 1H), 5.21 (d, J=4.8 Hz, 1H),3.82 (d, J=18 Hz, 1H), 3.66 (d, J=18 Hz, 1H), 3.21 (br, 2H), 2.99 (br,2H), 1.80 (br, 4H), 1.62 (s, 3H), 1.61 (s, 3H)

Example 43 Compound 43

Compound 43 (38.7 mg, 38%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-XXVIII.

¹H NMR (600 MHz, CD₃OD) δ 8.25 (s, 1H), 7.04 (s, 1H), 6.16 (m, 1H), 5.92(d, J=4.8 Hz, 1H), 5.22 (d, J=4.8 Hz, 1H), 4.87 (m, 2H), 3.98 (s, 2H),3.83 (d, J=18 Hz, 1H), 3.68 (d, J=18 Hz, 1H), 1.63 (s, 3H), 1.61 (s, 3H)

Example 44 Compound 44

Compound 44 (11.4 mg, 13%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-XXIX.

¹H NMR (600 MHz, CD₃OD) δ 8.11 (s, 1H), 7.00 (s, 1H), 6.76 (d, J=16.8Hz, 1H), 6.10 (d, J=15.6 Hz, 1H), 5.87 (br, 1H), 5.20 (br, 1H), 4.88 (m,2H), 3.78 (d, J=17.4 Hz, 1H), 3.67 (d, J=16.8 Hz, 1H), 3.11 (m, 4H),1.62 (s, 3H), 1.60 (s, 3H)

Example 45 Compound 45

Compound 45 (30 mg, 30%) was prepared by a method similar to Example 8by using Compound B-V

Compound A-XXX.

¹H NMR (600 MHz, CD₃OD) δ 8.23 (s, 1H), 7.03 (d, J=16.8 Hz, 1H), 6.98(s, 1H), 6.13 (m, 1H), 5.91 (d, J=5.4 Hz, 1H), 5.21 (d, J=4.8 Hz, 1H),4.90 (m, 2H), 3.81 (d, J=17.4 Hz, 1H), 3.66 (d, J=17.4 Hz, 1H), 3.29 (t,J=6 Hz, 2H), 2.93 (t, J=6 Hz, 2H), 1.62 (s, 3H), 1.60 (s, 3H)

Example 46 Compound 46

Compound 46 (20 mg, 18%) was prepared by a method similar to Example 8by using Compound B-XIII and Compound A-XXXI.

¹H NMR (600 MHz, DMSO d-₆) δ 9.48 (m, 1H), 8.93 (bs, 1H), 8.36 (s, 1H),7.97 (bs, 1H), 7.76 (bs, 1H), 7.75 (bs, 2H), 7.47 (bs, 1H), 7.42 (s,1H), 6.92 (d, J=16.2 Hz, 1H), 6.08 (m, 1H), 5.80 (m, 1H), 5.19 (d, J=4.8Hz, 1H), 4.82 (m, 2H), 3.80 (d, J=18.0 Hz, 1H), 3.55 (d, J=18.0 Hz, 1H),2.83 (m, 2H), 2.44 (m, 2H), 1.82 (m, 2H), 1.48 (s, 3H), 1.46 (s, 3H)

Example 47 Compound 47

Compound 47 (1 mg, 5%) was prepared by a method similar to Example 1 byusing Compound B-V and Compound A-XXXII.

¹H NMR (600 MHz, CD₃OD) δ=8.37 (s, 1H), 7.52 (s, 1H), 6.98 (s, 1H), 6.73(m, 1H), 6.01 (m, 2H), 5.21˜4.90 (m, 3H), 3.98 (m, 2H), 3.81 (m, 2H),1.58 (s, 3H), 1.56 (s, 3H)

Example 48 Compound 48

Compound 48 (52 mg, 58%) was prepared by a method similar to Example 1by using Compound B-VIII and Compound A-IX.

¹H NMR (600 MHz, CD₃OD) δ 8.89 (d, J=1.8 Hz, 1H), 8.72 (d, J=1.8 Hz,1H), 7.07 (s, 1H), 5.97 (d, J=5.4 Hz, 1H), 5.33˜4.8 (m, 3H), 3.71 (d,J=18.6 Hz, 1H), 3.41 (d, J=18.6 Hz, 1H), 3.26˜2.91 (m, 4H), 1.62 (s, 6H)

Example 49 Compound 49

Compound 49 (8 mg, 9%) was prepared by a method similar to Example 1 byusing Compound B-III and Compound A-I.

H NMR (600 MHz, DMSO d-₆) δ 9.54 (bs, 1H), 9.06 (bs, 1H), 8.44 (s, 1H),8.21 (bs, 3H), 7.56 (s, 1H), 6.86 (d, J=16.2 Hz, 1H), 6.29 (bs, 1H),6.05 (m, 1H), 5.76 (m, 1H), 5.13 (d, J=4.8 Hz, 1H), 4.85 (m, 2H), 3.66(d, J=16.8 Hz, 1H), 3.49 (d, J=17.4 Hz, 1H), 3.02 (m, 4H), 1.47 (s, 3H),1.46 (s, 3H)

Example 50 Compound 50

Compound 50 (3.5 mg, 7%) was prepared by a method similar to Example 1by using Compound B-V and Compound A-I.

¹H NMR (600 MHz, CD₃OD) δ 8.31 (s, 1H), 7.53 (s, 1H), 7.05 (m, 2H), 6.22(m, 1H), 5.90 (d, J=4.8 Hz, 1H), 5.19 (d, J=4.8 Hz, 1H), 3.80 (d, 17.4Hz 1H), 3.64 (d, J=17.4 Hz, 1H), 3.52 (t, J=6 Hz, 2H), 3.24 (t, J=6 Hz,2H) 1.61 (s, 3H), 1.59 (s, 3H)

Example 51 Compound 51

Compound 51 (48 mg, 60%) was prepared by a method similar to Example 8by using Compound B-V and Compound XXXI.

¹H NMR (600 MHz, CD₃OD) δ 8.33 (s, 1H), 7.58 (s, 1H), 7.11˜7.06 (m, 2H),6.27 (m, 1H), 5.92 (d, J=4.8 Hz, 1H), 5.23 (d, J=4.8 Hz, 1H), 4.93 (m,2H), 3.84 (d, J=18 Hz, 1H), 3.68 (d, J=18 Hz, 1H), 3.57 (br, 2H), 3.31(m, 2H), 2.76 (s, 3H), 1.64 (s, 3H), 1.63 (s, 3H)

Example 52 Compound 52

Compound 52 (18 mg, 30%) was prepared by a method similar to Example 8by using Compound B-VII and Compound A-I.

¹H NMR (600 MHz, CD₃OD) δ 8.31 (s, 1H), 7.54 (s, 1H), 7.06 (m, 2H), 6.22(m, 1H), 5.91 (d, J=5.4 Hz, 1H), 5.19 (d, J=5.4 Hz, 1H), 4.80 (m, 1H),3.80 (d, 17.4 Hz 1H), 3.65 (d, J=17.4 Hz, 1H), 3.50 (t, J=6 Hz, 2H),3.24 (t, J=6 Hz, 2H) 1.55 (d, J=6.6 Hz, 3H)

Example 53 Compound 53

Compound 53 (30 mg, 33%) was prepared by a method similar to Example 8by using Compound B-XII and Compound A-I.

¹H NMR (600 MHz, DMSO-d⁶) δ 9.53 (d, J=7.8 Hz, 1H), 9.11 (s, 1H), 8.47(s, 1H), 8.07 (s, 1H), 7.87 (s, 2H), 7.60 (s, 1H), 7.42 (s, 1H), 6.91(d, J=16.2 Hz, 1H), 6.26 (m, 1H), 6.02 (br, 1H), 5.84 (dd, J=4.8 Hz, 4.8Hz, 1H), 5.20 (d, J=4.8 Hz, 1H), 4.93 (m, 2H), 4.63 (q, J=7.2 Hz, 1H),3.81 (d, J=18 Hz, 1H), 3.58 (d, J=18 Hz, 1H), 3.32 (m, 2H), 3.06 (br,2H), 1.43 (d, J=7.2 Hz, 3H)

Example 54 Compound 54

Compound 54 (29 mg, 30%) was prepared by a method similar to Example 8by using Compound B-XIII and Compound A-I.

¹H NMR (600 MHz, DMSO d-₆) δ 11.77 (t, J=5.4 Hz, 1H), 11.04 (bs, 1H),10.91 (bs, 1H), 9.45 (d, J=8.4 Hz, 1H), 9.00 (bs, 1H), 8.42 (s, 1H),8.09 (bs, 1H), 7.82 (s, 1H), 7.70 (s, 1H), 7.57 (s, 1H), 7.42 (s, 1H),6.90 (d, J=16.2 Hz, 1H), 6.29 (m, 1H), 6.05 (t, J=4.8 Hz, 1H), 5.82 (dd,J₁=8.4 Hz, J₂=5.4 Hz, 1H), 5.20 (d, J=5.4 Hz, 1H), 4.88 (m, 2H), 3.83(d, J=18.0 Hz, 1H), 3.58 (m, 3H), 3.29 (m, 2H), 1.47 (s, 3H), 1.45 (s,3H)

Example 55 Compound 55

Compound 55 (26 mg, 29%) was prepared by a method similar to Example 8by using Compound B-XI and Compound A-I.

¹H NMR (600 MHz, DMSO d-₆) δ 9.54 (bs, 1H), 9.06 (bs, 1H), 8.44 (s, 1H),8.21 (bs, 3H), 7.56 (s, 1H), 6.86 (d, J=16.2 Hz, 1H), 6.29 (bs, 1H),6.05 (m, 1H), 5.76 (m, 1H), 5.13 (d, J=4.8 Hz, 1H), 4.85 (m, 2H), 3.66(d, J=16.8 Hz, 1H), 3.49 (d, J=17.4 Hz, 1H), 3.02 (m, 4H), 1.47 (s, 3H),1.46 (s, 3H)

Example 56 Compound 56

Compound 56 (38 mg, 32%) was prepared by a method similar to Example 8by using Compound B-XI and Compound A-XVII.

¹H NMR (600 MHz, DMSO d-₆) δ 9.48 (d, J=9.0 Hz, 1H), 9.04 (bs, 1H), 8.42(s, 1H), 8.21 (bs, 3H), 7.57 (s, 1H), 6.91 (d, J=16.2 Hz, 1H), 6.14 (m,2H), 5.76 (bs, 1H), 5.14 (m, 1H), 4.84 (bs, 1H), 3.53 (m, 2H), 3.21 (m,2H), 2.90 (m, 2H), 1.88 (m, 2H), 1.46 (s, 3H), 1.46 (s, 3H)

Example 57 Compound 57

Compound 57 (30 mg, 25%) was prepared by a method similar to Example 8by using Compound B-XIII and Compound A-XVI.

¹H NMR (600 MHz, DMSO d-₆) δ 9.41 (m, 1H), 9.04 (bs, 1H), 8.42 (s, 1H),8.22 (bs, 2H), 7.58 (bs, 1H), 7.42 (s, 1H), 6.91 (d, J=15.6 Hz, 1H),6.13 (m, 2H), 5.73 (m, 1H), 5.13 (m, 1H), 4.85 (m, 2H), 3.52 (m, 2H),3.22 (bs, 2H), 2.89 (bs, 2H), 1.87 (m, 2H), 1.48 (s, 3H), 1.46 (s, 3H)

Example 58 Compound 58

Compound 58 (9 mg, 24%) was prepared by a method similar to Example 8 byusing Compound B-XIII and Compound A-XXVII.

¹H NMR (600 MHz, CD₃OD) δ 8.27 (s, 1H), 7.41 (s, 1H), 7.10 (d, J=15.6Hz, 1H), 6.27 (m, 1H), 5.90 (d, J=4.8 Hz, 1H), 5.21 (d, J=4.8 Hz, 1H),4.91 (m, 2H), 3.85 (d, J=18.6 Hz, 1H), 3.66 (d, J=18 Hz, 1H), 3.21 (br,2H), 3.00 (br, 2H), 1.80 (br, 4H), 1.60 (s, 3H), 1.59 (s, 3H)

Example 59 Compound 59

Compound 59 (47 mg, 49%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-XXXIII.

¹H NMR (600 MHz, CD₃OD) δ 8.65 (s, 1H), 8.60 (s, 1H), 7.19 (d, J=16.2Hz, 1H), 6.97 (s, 1H), 6.20 (m, 1H), 5.92 (d, J=4.8 Hz, 1H), 5.21 (d,J=4.8 Hz, 1H), 4.93˜4.83 (m, 2H), 3.98 (s, 2H), 3.80 (d, J=18 Hz, 1H),3.66 (d, J=18 Hz, 1H), 1.62 (s, 3H), 1.6 (s, 3H)

Example 60 Compound 60

Compound 60 (49 mg, 53%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-IX.

¹H NMR (600 MHz, CD₃OD) δ 8.63 (s, 1H), 8.62 (s, 1H), 7.17 (d, J=16.2Hz, 1H), 6.97 (s, 1H), 6.19 (m, 1H), 5.92 (d, J=4.8 Hz, 1H), 5.22 (d,J=5.4 Hz, 1H), 4.92˜4.8 (m, 2H), 3.80 (d, J=18 Hz, 1H), 3.66 (d, J=17.4Hz, 1H), 3.28 (m, 2H), 2.91 (t, J=6 Hz, 2H), 1.62 (s, 3H), 1.60 (s, 3H)

Example 61 Compound 61

Compound 61 (6.8 mg, 10%) was prepared by a method similar to Example 8by using Compound B-XI and Compound A-IX.

¹H NMR (600 MHz, CD3OD) δ 8.62 (s, 1H), 8.60 (s, 1H), 7.23 (d, J=15.6Hz, 1H), 6.22 (m, 1H), 5.92 (d, J=4.8 Hz, 1H), 5.19 (d, J=4.8 Hz, 1H),4.91 (m, 2H), 3.80 (d, 17.4 Hz 1H), 3.64 (d, J=17.4, 1H), 3.28 (m, 2H),2.90 (t, J=6 Hz, 2H), 1.60 (s, 3H), 1.58 (s, 3H)

Example 62 Compound 62

Compound 62 (11 mg, 13%) was prepared by a method similar to Example 8by using Compound B-XIII and Compound A-IX.

¹H NMR (600 MHz, CD₃OD) δ 8.45 (s, 1H), 8.43 (s, 1H), 7.09 (d, J=16.2Hz, 1H), 6.12 (m, 1H), 5.87 (d, J=4.8 Hz, 1H), 5.17 (d, J=5.4 Hz, 1H),4.89 (m, 2H), 3.74 (d, J=17.4 Hz 1H), 3.61 (d, J=17.4 Hz, 1H), 3.30 (m,2H), 2.90 (m, 2H), 1.59 (s, 3H), 1.57 (s, 3H)

Example 63 Compound 63

Compound 63 (46 mg, 57%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-XXVI.

¹H NMR (600 MHz, CD₃OD) δ 8.66 (d, J=1.8 Hz, 1H), 8.61 (d, J=1.8 Hz),7.18 (d, 15.6 Hz, 1H), 6.96 (s, 1H), 6.19 (m, 1H), 5.92 (d, J=4.8 Hz,1H), 5.22 (d, J=5.4 Hz, 1H), 4.91 (m, 2H), 3.81 (d, J=18 Hz, 1H), 3.66(d, J=18 Hz, 1H), 3.04 (t, J=7.2 Hz, 2H), 2.66 (t, J=6.6 Hz, 2H), 2.04(m, 2H), 1.62 (s, 3H), 1.60 (s, 3H)

Example 64 Compound 64

Compound 64 (12 mg, 14%) was prepared by a method similar to Example 8by using Compound B-XIII and Compound A-XXVI.

¹H NMR (600 MHz, CD₃OD) δ 8.69 (s, 1H), 8.59 (s, 1H), 7.20 (d, J=16.2Hz, 1H), 6.24 (m, 1H), 5.92 (d, J=4.8 Hz, 1H), 5.21 (d, J=4.8 Hz, 1H),4.93 (m, 2H), 3.84 (d, J=18 Hz, 1H), 3.67 (d, J=18 Hz, 1H), 3.03 (br,2H), 2.66 (br, 2H), 2.03 (br, 2H), 1.60 (s, 3H), 1.59 (s, 3H)

Example 65 Compound 65

Compound 65 (39 mg, 48%) was prepared by a method similar to Example 8by using Compound B-XI and Compound A-XXVI.

¹H NMR (600 MHz, CD₃OD) δ 8.65 (s, 1H), 8.61 (s, 1H), 7.19 (d, J=15.6Hz, 1H), 6.22 (m, 1H), 5.93 (d, J=4.8 Hz, 1H), 5.21 (d, J=4.8 Hz, 1H),4.92 (m, 2H), 3.82 (d, J=18 Hz, 1H), 3.66 (d, J=17.4 Hz, 1H), 3.04 (t,J=7.2 Hz, 2H), 2.66 (t, J=6.6 Hz, 2H), 2.04 (m, 2H), 1.62 (s, 3H), 1.60(s, 3H)

Example 66 Compound 66

Compound 66 (30 mg, 32%) was prepared by a method similar to Example 8by using Compound B-V and Compound A-XXX.

¹H NMR (600 MHz, CD₃OD) δ 8.23 (s, 1H), 7.03 (d, J=16.8 Hz, 1H), 6.98(s, 1H), 6.13 (m, 1H), 5.91 (d, J=5.4 Hz, 1H), 5.21 (d, J=4.8 Hz, 1H),4.90 (m, 2H), 3.81 (d, J=17.4 Hz, 1H), 3.66 (d, J=17.4 Hz, 1H), 3.29 (t,J=6 Hz, 2H), 2.93 (t, J=6 Hz, 2H), 1.62 (s, 3H), 1.60 (s, 3H)

Example 67 Compound 67

Compound 67 (20 mg, 18%) was prepared by a method similar to Example 8by using Compound B-XIII and Compound A-XXXI.

¹H NMR (600 MHz, DMSO d-₆) δ 9.48 (m, 1H), 8.93 (bs, 1H), 8.36 (s, 1H),7.97 (bs, 1H), 7.76 (bs, 1H), 7.75 (bs, 2H), 7.47 (bs, 1H), 7.42 (s,1H), 6.92 (d, J=16.2 Hz, 1H), 6.08 (m, 1H), 5.80 (m, 1H), 5.19 (d, J=4.8Hz, 1H), 4.82 (m, 2H), 3.80 (d, J=18.0 Hz, 1H), 3.55 (d, J=18.0 Hz, 1H),2.83 (m, 2H), 2.44 (m, 2H), 1.82 (m, 2H), 1.48 (s, 3H), 1.46 (s, 3H)

Example 68 Compound 68

B-IX Compound (170 mg, 0.182 mmol) and 4,5-diaminopyrimidine (20 mg,0.182 mmol) were dissolved in N,N-dimethylformamide (1.5 mL) and sodiumbromide (37.5 mg, 0.364 mmol) was added. The resulting solution wasstirred for 9 hours at room temperature. Potassium iodide (211 mg, 1.27mmol) and N,N-dimethylformamide (1 mL) were added. At −40° C. acetylchloride (71 mg, 0.91 mmol) was added while stirring, and stirred for 5minutes at the same temperature and further stirred for 1 hour at 0° C.An aqueous solution of sodium thiosulfate pentahydrate dissolved insaline (5 mL) was added to the resulting solution at 0° C. Thethus-obtained solid was dissolved in methylene chloride (15 mL) andapplied to column chromatography (MC:MeOH=50:1˜10:1) to yield aquaternary salt compound (78 mg (42%)). The quaternary salt compound (78mg, 0.075 mmol) was dissolved in methylene chloride (0.5 mL).Triethylsilane (0.5 mL) and trifluoroacetic acid (1.5 mL) weresequentially added. The resulting solution was stirred for 4 hours atroom temperature. Isopropylether (25 mL) was added to the resultingsolution, creating a solid. The solid was filtered under reducedpressure to yield Compound 68 (50 mg, 99%) (In the middle of thereaction, due to acetyl chloride, acetylation occurred at 5 position of4,5-diaminopyrimidine).

¹H NMR (600 MHz, CD₃OD) δ 8.79 (d, J=1.8 Hz, 1H), 8.70 (d, J=1.8 Hz,1H), 7.08 (s, 1H), 6.00 (d, J=4.8 Hz, 1H) 5.32 (d, J=14.4 Hz, 1H), 5.27(d, J=4.8 Hz, 1H), 4.8 (d, J=14.4 Hz, 1H), 3.71 (d, J=18.6 Hz, 1H), 3.44(d, J=18 Hz, 1H), 1.60 (d, J=1.2 Hz, 6H)

Test Example 1 In Vitro Antibacterial Activity Test

To evaluate antibacterial activity of each of the compounds of Examples1 to 68, in vitro antibacterial activity test was performed. The invitro antibacterial activity was evaluated by measuring MIC₉₀ (ug/mL) ofeach of the compounds of Examples 1 to 68, which is defined as thelowest concentration of an antibiotic that will inhibit the visiblegrowth of 90% of microorganisms after incubation as compared with acontrol group to which the antibiotic is not treated. MIC_(H) valueswere measured by the broth microdilution method developed by theClinical and Laboratory Standards Institute (CLSI) (see CLSI M7-A5,Methods for Dilution Antimicrobial Susceptibility Test for Bacteria thatGrow Aerobically-Fifth Edition (2000): CLSI, Villanova, Pa.).

Ceftazidime (CAZ) represented by Formula B, CXA-101 represented byFormula C, and Doripenem represented by Formula D were used ascomparison compounds. The test results are shown in Tables 1 and 2.

Ceftazidime represented by Formula B is a third-generation cephalosporinantibiotic and is widely used against Pseudomonas aeruginosa. CXA-101represented by Formula C is a cephalosporin antibiotic which is in phase2 clinical trials by Cubist Pharmaceuticals, Inc. Doripenem representedby Formula D belongs to a subgroup of cabapenems and is one of theantibiotics that are most widely used to treat drug-resistantGram-negative infection.

1) Test Bacteria

In vitro antibacterial activity was measured with respect to thefollowing 19 clinical isolates: M. catarrhalis; P. aeruginosa (5strains); K. pneumoniae (6 strains); A. baumannii (3 strains); E. coli(2 strains); A. calcoaceticus; and E. cloacae. Table 1 shows the result.

2) Preparation of Test Compositions

Test compounds each (the cephalosporin derivative compounds prepared inExamples 1 to 68) were dissolved in DMSO at the concentration of 10,240ug/mL, were diluted by two fold with DMSO, and then were diluted bytwenty fold with sterilized distilled water. The final concentration inthe antibacterial activity test was in the range of 0.0626 ug/mL to 128ug/mL, and the final concentration of DMSO used as an adjuvant was 2.5%(V/V).

TABLE 1 Antibacterial Activity of Compounds of Formula I (MIC₉₀, ug/mL)Test Cephalosporin derivative compounds according to the presentinvention Bacteria 1 2 3 4 5 6 7 8 CAZ 1 M. catarrhalis 1 1 2 0.25 0.252 2 0.5 <0.0625 2 P. aeruginosa 0.25 2 32 0.5 1 0.5 0.5 1 1 3 P.aeruginosa 1 16 0.5 0.5 4 2 1 4 32 4 P. aeruginosa 0.5 4 1 0.5 1 0.50.125 1 64 5 P. aeruginosa 0.25 4 0.125 1 1 0.5 0.125 0.5 4 6 P.aeruginosa 0.25 8 0.25 1 0.25 0.5 0.25 1 16 7 K. pneumoniae 2 4 1 2 2 42 1 1 8 K. pneumoniae 4 0.5 <0.0625 2 <0.0625 0.5 16 9 K. pneumoniae0.125 0.5 1 0.5 4 <0.0625 32 10 K. pneumoniae 128 4 2 8 16 32 64 11 K.pneumoniae 0.5 1 0.5 0.125 <0.0625 0.25 32 12 K. pneumoniae 128 64 64 328 16 32 8 64 13 A. baumannii 4 4 2 64 16 32 4 8 4 14 A. baumannii16 >128 >128 8 >128 >128 8 >128 >64 15 A. baumannii 32 >128 1284 >128 >128 16 >128 >64 16 A. calcoacetius 0.5 2 0.25 2 2 2 1 2 1 17 E.coli <0.0625 0.25 <0.0625 0.125 <0.0625 0.25 <0.0625 0.0625 0.5 18 E.coli 2 4 1 0.5 0.5 4 0.125 0.5 8 19 E. cloacae 2 >128 2 1 0.5 2 1 32 32Test Cephalosporin derivative compounds according to the presentinvention Bacteria 9 10 11 12 13 14 15 16 17 1 M. catarrhalis 0.5 0.50.25 1 2 0.25 0.25 1 0.125 2 P. aeruginosa 0.25 4 1 1 2 1 0.5 1 2 3 P.aeruginosa 0.25 4 4 2 8 2 1 2 2 4 P. aeruginosa 0.25 2 1 8 8 16 0.1250.5 8 5 P. aeruginosa <0.0625 1 0.5 1 2 0.25 0.5 0.25 8 6 P. aeruginosa0.25 1 1 1 4 0.25 0.25 2 2 7 K. pneumoniae 0.125 2 0.5 2 2 2 8 8 4 8 K.pneumoniae 1 0.25 <0.0625 0.25 1 2 4 128 1 9 K. pneumoniae 0.125 <0.0625<0.0625 0.25 1 0.5 0.25 2 4 10 K. pneumoniae >128 32 >128 64 64 6464 >128 32 11 K. pneumoniae 0.25 2 <0.0625 <0.0625 1 0.5 2 32 1 12 K.pneumoniae 64 8 8 8 8 8 128 128 32 13 A. baumannii 0.5 8 4 4 8 2 128 216 14 A. baumannii >128 >128 >128 >128 >128 >128 8 >128 4 15 A.baumannii >128 >128 >128 >128 128 8 2 32 2 16 A. calcoacetius 0.5 2 1 12 0.25 0.5 0.25 0.5 17 E. coli <0.0625 0.125 0.125 0.25 1 0.25 2 1 1 18E. coli 1 0.5 0.125 0.25 2 1 16 4 4 19 E. cloacae 2 0.5 64 4 8 64 32 162 Test Cephalosporin derivative compounds according to the presentinvention Bacteria 18 19 20 21 22 23 24 25 26 1 M. catarrhalis 0.5 1 0.50.125 0.5 0.25 <0.0625 <0.0625 <0.0625 2 P. aeruginosa 1 2 4 4 2 1 0.250.25 0.25 3 P. aeruginosa 2 2 1 4 2 2 0.5 0.5 0.5 4 P. aeruginosa 0.250.5 2 32 1 0.25 <0.0625 <0.0625 <0.0625 5 P. aeruginosa 2 0.5 4 8 2 0.50.25 <0.0625 <0.0625 6 P. aeruginosa 0.25 2 1 2 0.5 1 0.25 0.25 0.25 7K. pneumoniae 16 16 8 4 16 1 0.5 0.25 0.5 8 K. pneumoniae 8 64 2 2 8<0.0625 <0.0625 <0.0625 0.125 9 K. pneumoniae 4 1 0.5 2 1 <0.0625<0.0625 <0.0625 <0.0625 10 K. pneumoniae 64 >128 16 8 64 64 2 >128 0.511 K. pneumoniae 4 16 1 2 2 <0.0625 0.125 <0.0625 <0.0625 12 K.pneumoniae 128 128 64 16 128 4 4 16 4 13 A. baumannii 128 4 16 16 128 42 0.5 2 14 A. baumannii 8 16 32 8 16 >128 32 128 16 15 A. baumannii 2 82 2 2 >128 16 8 2 16 A. calcoacetius 1 0.5 1 1 0.5 1 0.5 0.25 0.5 17 E.coli 2 1 0.25 0.5 2 <0.0625 <0.0625 <0.0625 <0.0625 18 E. coli 16 16 8 432 <0.0625 <0.0625 <0.0625 <0.0625 19 E. cloacae 64 128 8 16 32 2 0.5 10.125 Test Cephalosporin derivative compounds according to the presentinvention Bacteria 27 28 29 30 31 32 33 34 35 47 1 M. catarrhalis 0.25 81 0.25 2 1 <0.0625 0.25 2 P. aeruginosa 1 32 2 1 1 2 64 32 2 8 3 P.aeruginosa 4 16 8 0.5 4 4 128 1 2 128 4 P. aeruginosa 2 8 2 2 1 4 >128 20.25 64 5 P. aeruginosa 2 16 4 0.5 2 4 128 0.5 1 32 6 P. aeruginosa 1 44 0.5 1 0.5 >128 0.5 1 128 7 K. pneumoniae 2 8 8 0.5 8 1 16 0.25 2 4 8K. pneumoniae 9 K. pneumoniae 10 K. pneumoniae 11 K. pneumoniae 12 K.pneumoniae 32 64 16 16 32 64 128 128 16 32 13 A. baumannii 16 8 8 1 4 1128 2 4 16 14 A. baumannii >128 >128 32 >128 32 32 >128 128 4 >128 15 A.baumannii >128 16 32 128 64 16 >128 128 2 128 16 A. calcoacetius 4 1 4 11 0.5 128 0.5 0.5 8 17 E. coli 1 >128 2 0.125 0.25 <0.0625 16 <0.0625<0.0625 >128 18 E. coli 2 8 2 2 8 8 64 1 1 16 19 E. cloacae 16 128 8 416 128 >128 16 0.5 128 Test Cephalosporin derivative compounds accordingto the present invention Bacteria 36 37 38 39 40 41 42 43 44 1 M.catarrhalis 0.125 <0.0625 <0.0625 <0.0625 2 P. aeruginosa 8 4 32 8 4 4 24 8 3 P. aeruginosa 64 8 128 128 16 16 32 16 32 4 P. aeruginosa >12816 >128 128 8 8 16 32 16 5 P. aeruginosa 16 4 64 32 8 4 8 16 16 6 P.aeruginosa 4 32 128 >128 >128 128 7 K. pneumoniae 0.5 8 4 1 16 4 0.5 0.50.5 8 K. pneumoniae 2 8 1 1 9 K. pneumoniae 2 0.5 0.5 0.5 10 K.pneumoniae 8 >128 32 32 11 K. pneumoniae 2 1 0.5 1 12 K. pneumoniae 1616 64 8 8 2 2 1 1 13 A. baumannii 64 4 128 8 2 32 2 32 16 14 A.baumannii >128 128 >128 >128 128 32 64 >128 >128 15 A. baumannii >12864 >128 >128 128 32 64 >128 >128 16 A. calcoacetius 2 1 16 8 0.25 1 0.54 1 17 E. coli 0.5 4 8 1 4 1 0.125 0.25 0.25 18 E. coli 16 4 16 16 8 10.5 0.5 0.5 19 E. cloacae >128 16 >128 128 32 4 16 0.5 0.5 TestCephalosporin derivative compounds according to the present inventionBacteria 45 46 48 49 50 51 52 53 54 1 M. catarrhalis <0.0625 <0.0625<0.0625 <0.0625 <0.0625 <0.0625 <0.0625 2 P. aeruginosa 4 4 8 2 2 2 0.51 2 3 P. aeruginosa 16 32 16 16 8 32 4 8 16 4 P. aeruginosa 16 16 64 8 832 16 8 4 5 P. aeruginosa 8 16 8 4 4 4 2 8 4 6 P.aeruginosa >128 >128 >128 64 64 32 32 64 7 K. pneumoniae 0.5 2 4 8 0.250.5 <0.0625 1 1 8 K. pneumoniae 0.5 8 0.5 0.5 4 1 1 9 K. pneumoniae 0.54 0.25 0.25 1 1 0.5 10 K. pneumoniae 16 8 16 32 128 2 2 11 K. pneumoniae0.5 2 0.125 0.5 4 2 1 12 K. pneumoniae 1 4 16 32 0.25 0.5 8 2 1 13 A.baumannii 16 64 8 8 4 8 1 8 4 14 A. baumannii >128 64 128 6464 >128 >128 64 128 15 A. baumannii >128 64 128 64 128 128 >128 64 32 16A. calcoacetius 2 4 0.5 2 0.5 0.5 1 2 1 17 E. coli 0.25 0.5 2 4 0.1250.125 <0.0625 0.25 0.25 18 E. coli 0.25 2 2 8 <0.0625 0.125 <0.0625 10.5 19 E. cloacae 0.25 32 128 32 8 32 8 4 4 Test Cephalosporinderivative compounds according to the present invention Bacteria 55 5657 58 59 60 61 62 63 1 M. catarrhalis <0.0625 <0.0625 <0.0625 <0.0625<0.0625 <0.0625 <0.0625 <0.0625 <0.0625 2 P. aeruginosa 1 2 0.5 8 4 2 24 2 3 P. aeruginosa 8 16 16 32 32 16 32 16 16 4 P. aeruginosa 4 8 8 3264 32 32 8 32 5 P. aeruginosa 1 8 4 16 16 8 2 8 4 6 P. aeruginosa >12864 >128 >128 128 64 >128 64 128 7 K. pneumoniae <0.0625 2 0.25 2 0.250.25 0.25 1 0.25 8 K. pneumoniae 2 1 1 8 2 1 16 1 1 9 K. pneumoniae0.125 1 0.25 2 1 0.5 1 0.5 0.5 10 K. pneumoniae 128 2 128 8 128 64 >1281 32 11 K. pneumoniae 0.25 1 0.5 4 1 0.5 2 1 1 12 K. pneumoniae 0.5 2 14 2 0.5 4 1 1 13 A. baumannii 0.5 4 1 32 16 8 128 16 8 14 A. baumannii64 32 64 32 >128 >128 128 32 128 15 A. baumannii 64 16 64 64 >128 128128 32 128 16 A. calcoacetius 0.125 1 0.5 4 2 1 1 1 1 17 E. coli <0.06250.5 0.125 0.5 0.25 0.125 0.125 0.5 0.125 18 E. coli <0.0625 1 0.125 20.5 0.25 0.125 0.5 0.25 19 E. cloacae 8 4 8 8 0.5 0.5 16 4 16 TestCephalosporin derivative compounds according to the present inventionBacteria 64 65 66 67 68 1 M. catarrhalis <0.0625 <0.0625 <0.0625 <0.06252 P. aeruginosa 4 2 4 4 4 3 P. aeruginosa 32 32 16 32 16 4 P. aeruginosa16 32 16 16 128 5 P. aeruginosa 8 4 8 16 4 6 P. aeruginosa128 >128 >128 >128 7 K. pneumoniae 2 0.25 0.5 2 1 8 K. pneumoniae 2 160.5 8 9 K. pneumoniae 1 1 0.5 4 10 K. pneumoniae 8 >128 16 8 11 K.pneumoniae 2 2 0.5 2 12 K. pneumoniae 2 2 1 4 32 13 A. baumannii 16 6416 64 8 14 A. baumannii 32 64 >128 64 128 15 A. baumannii 32 64 >128 64128 16 A. calcoacetius 1 0.5 2 4 1 17 E. coli 0.5 <0.0625 0.25 0.5 0.518 E. coli 1 0.125 0.25 2 8 19 E. cloacae 8 32 0.25 32 128 1: M.catarrhalis 2524 2: P. aeruginosa 1912E, 3: P. aeruginosa 6065Y, 4: P.aeruginosa 37, 5: P. aeruginosa 40, 6: P. aeruginosa 43, 7: K.pneumoniae 2011E, 8: K. pneumoniae β9, 9: K. pneumoniae β10, 10: K.pneumoniae β11, 11: K. pneumoniae β13, 12: K. pneumoniae β14, 13: A.baumannii 46, 14: A. baumannii 49, 15: A. baumannii 52, 16: A.calcoaceticus ATCC15473, 17: E. coli AG100, 18: E. coli β4, 19: E.cloacae β19

As shown in Table 1, the antibacterial activities of the compounds witha siderophore group were significantly greater than those of thecompounds without a siderophore group. The antibacterial activitiesvaried depending on the kind of the siderophore group and the site atwhich the group is introduced. The antibacterial activity was greatlyaffected by the site to which the group is introduced.

For the Compounds 4, 8, 11, and 26, which exhibited antibacterialactivity significantly improved by introduction of a siderophore group,antibacterial activity was then evaluated with regard to therepresentative drug-resistance Gram-negative bacteria: P. aeruginosa (16clinical isolates), K. pneumoniae (38 clinical isolates), A. baumannii(6 clinical isolates). The antibacterial activity of the four compoundswas compared with that of the comparison compounds, which is shown inTable 2. The numerical values in Table 2 represent the number ofclinical isolates that showed the respective MIC values.

TABLE 2 Antibacterial Activity on Representative Bacteria (MIC₉₀, ug/mL)MIC(ug/mL) >16 16 8 4 2 1 0.5 0.25 0.125 0.0625 <0.0625 P. aeruginosa (N= 16) Ceftazidime 11 1 1 2 1 CXA-101 7 1 4 4 Doripenem 9 5 1 1 Compound4 3 1 1 2 1 2 5 1 Compound 8 3 1 3 2 2 2 1 2 Compound 11 2 1 2 3 2 2 3 1Compound 26 3 1 3 2 1 3 2 1 A. baumanni (N = 6) Ceftazidime 6 CXA-101 6Doripenem 5 1 Compound 4 3 2 1 Compound 8 5 1 Compound 11 5 1 Compound26 3 2 1 K. pneumomiae (N = 38) Ceftazidime 32 3 1 2 CXA-101 35 1 1 1Doripenem 2 4 5 6 5 3 6 1 6 Compound 4 26 2 3 2 3 1 1 Compound 8 35 1 11 Compound 11 35 1 1 1 Compound 26 22 4 4 1 3 2 1 1

As shown in Table 2, the cephalosporin derivatives according to thepresent invention which have a siderophore group exhibited superiorantibacterial spectrum compared with the comparison compounds,Ceftazidime, CXA-101, and Doripenem, and they exhibited excellentactivity against, in particular, P. aeruginosa, suggesting that theyhave a great potential to be used to treat drug-resistance Gram-negativeinfection.

Test Example 2 In Vivo Pharmaceutical Efficacy Test

Pharmaceutical efficacy of the cephalosporin derivative compoundsaccording to the present invention was evaluated in a whole-bodyinfected mouse model. Tables 3 and 4 show survival rate and ED₅₀ valuesfor the two compounds with excellent antibacterial activity with regardto an infected mouse by a drug-sensitive bacteria and a drug-resistantbacteria.

Test animals: 3 week old male ICR mouse, weight 18˜22 g; 5 mice/group.Lab conditions: Temperature of 23±2° C.; humidity of 55±20%.Administration method: Inducing whole-body infection by bacterialsolution followed by subcutaneous injection (0.2 mL) after 1 hour and 4hours.Test method: Cultured bacteria were diluted with 0.9% NaCl to prepare abacterial solution having a concentration that is 5 to 10 times ofminimal inhibitory concentration (MIC). 0.5 mL of bacterial solution wasinjected through abdominal cavity to induce whole-body infection. Testcompounds were administered in four different amounts, which weredesigned considering in vitro MIC values of the test bacteria. After 1hour and 4 hours, the four different amounts of the test compounds weresubcutaneously administered. Survival rates were measured for four daysand ED₅₀ values were calculated according to the Probit method.

TABLE 3 Efficacy on Infected Mouse by Ceftazidime- sensitive P.aeruginosa Ceftazidime Compound 8 Administered amount (mg/kg) 20 10 52.5 20 10 5 2.5 Survival rate 100% 100% 80% 40% 100% 100% 80% 60% ED₅₀(mg/kg) 2.50 1.25 (95% sd) (0.74~8.49) (0.09~17.5) Infecting bacteria:P. aeruginosa 1912E (2 × 10⁶ CFU/mouse)

TABLE 4 Efficacy on Infected Mouse by Ceftazidime-resistant P.aeruginosa Ceftazidime Doripenem Compound 4 Administered amount (mg/kg)40 20 10 5 40 20 10 5 40 20 10 5 Survival rate 20% 20% 20% 0% 80% 20%20% 0% 100% 80% 40% 40% ED₅₀ (mg/kg) 640 28 10 (95% sd) (0.12~3290000)(13.8~57.8) (3.97~25.2) Infecting bacteria: P. aeruginosa R1023 (2 × 10⁶CFU/mouse)

For the whole-body infected mice by the drug-sensitive bacteria,pharmaceutical efficacy of Compound 8 was similar to that of thecomparison compound, Ceftazidime. For the whole-body infected mice bythe drug-resistant bacteria, the pharmaceutical efficacy of Compound 4was much greater than that of Ceftazidime and greater than that ofDoripenem that is known to be the best treating agent against P.aeruginosa.

As shown in the results, the compounds according to the presentinvention showed superior pharmaceutical efficacy in in vitro and invivo and maintained it even in case where siderophore is introduced. Incase of catechol, a typical siderophore, in vivo pharmaceutical efficacywas reported to be sharply decreased by catechol O-methyl transferase(COMT). On the other hand, the compounds according to the presentinvention did not show such a decrease. It suggests that the presentcompounds are capable of being used as a treating agent againstdrug-resistant bacteria that are known to be difficult to be treated.

Test Example 3 Pharmacokinetics Study

For the present compounds with superior pharmaceutical efficacy, PKvalues were evaluated in a rat model. Table 5 shows the results of tworepresentative compounds.

Test animals: 9 week old SD rat, weight 290˜310 g; 3 rats/sample timepointLab conditions: Temperature of 21±2° C.; humidity of 50±20%Administration method: Injecting test compound solution through tailvein (IV)Test method: Blood samples were taken from jugular vein at apredetermined time period for 24 hours after administration, plasma wasseparated, and quantified by using LC-MS/MS.

TABLE 5 Pharmacokinetic Test Results Compound 4 Compound 8 Rat (10mg/kg) single IV Rat (10 mg/kg) single IV AUC (mg*h/l) 40.28 54.60AUC_(norm) (kg*h/l) 4.03 5.46 CL (l/h/kg) 0.25 0.18 V_(ss) (l/kg) 0.200.09 C_(max) (mg/l) 75.80 196.13 C_(max, norm) (kg/l) 7.58 19.61 t_(max)(h) 0.02 0.02 t_(1/2) (h) 0.62 0.87

As shown in the Table above, the cephalosporin derivatives according tothe present invention maintained higher concentration in blood andexhibited an excellent pharmacokinetic profile, thereby being able to beused as a promising antibiotic.

The foregoing descriptions of specific exemplary embodiments of thepresent invention have been presented for purposes of illustration anddescription. They are not intended to be exhaustive or to limit theinvention to the precise forms disclosed, and obviously manymodifications and variations are possible in light of the aboveteachings. The exemplary embodiments were chosen and described in orderto explain certain principles of the invention and their practicalapplication, to thereby enable others skilled in the art to make andutilize various exemplary embodiments of the present invention, as wellas various alternatives and modifications thereof. It is intended thatthe scope of the invention be defined by the Claims appended hereto andtheir equivalents.

1. A novel cephalosporin derivative represented by Chemical Formula 1,an in vivo hydrolysable ester thereof, an in vivo hydrolysable phosphateester thereof, an isomer thereof, or a pharmaceutically acceptable saltthereof:

wherein X is CR, N, or C substituted by Cl (C—Cl); Y is C₁-C₂ alkyl,CH(CH₃)CO₂H, or C(CH₃)₂CO₂H; L is CH₂ or CH═CHCH₂; R₁ is NH₂, NHR₁₁ orNH(CH₂)_(m)NR₁₁R₁₂; R₂ is NHR₂₁, NH(CH₂)_(n)COOH, NH(CH₂)_(n)NR₂₁R₂₂, orNHC(═O)(CH₂)_(n)NR₂₁R₂₂; and R₃ is hydrogen or NH₂, in which R ishydrogen or C₁-C₃ alkyl; R₁₁ and R₂₁ are independently hydrogen, C₁-C₃alkyl, or a group selected from the group consisting of:

R₁₂ and R₂₂ are independently hydrogen or C₁-C₂ alkyl; and m and n areindependently an integer of 1 to
 6. 2. The novel cephalosporinderivative of claim 1, which is represented by Chemical Formula 2, an invivo hydrolysable ester thereof, an in vivo hydrolysable phosphate esterthereof, an isomer thereof, or a pharmaceutically acceptable saltthereof:

wherein X is CR, N, or C substituted by Cl (C—Cl); Y is C₁-C₂ alkyl,CH(CH₃)CO₂H, or C(CH₃)₂CO₂H; L is CH₂ or CH═CHCH₂; R₁ is NH₂, NHR₁₁ orNH(CH₂)_(m)NR₁₁R₁₂; and R₂ is NHR₂₁, NH(CH₂)_(n)NR₂₁R₂₂, orNHC(═O)(CH₂)_(n)NR₂₁R₂₂, in which R is hydrogen or C₁-C₃ alkyl; R₁₁ andR₂₁ are independently hydrogen, C₁-C₃ alkyl, or a group selected fromthe group consisting of:

R₁₂ and R₂₂ are independently hydrogen or C₁-C₂ alkyl; and m and n areindependently an integer of 1 to
 6. 3. The novel cephalosporinderivative of claim 2, an in vivo hydrolysable ester thereof, an in vivohydrolysable phosphate ester thereof, an isomer thereof, or apharmaceutically acceptable salt thereof, wherein X is CR, N, or Csubstituted by Cl (C—Cl); Y is CH(CH₃)CO₂H or C(CH₃)₂CO₂H; L is CH₂ orCH═CHCH₂; R₁ is NH₂ or NH(CH₂)_(m)NH₂; R₂ is NHR₂₁, NH(CH₂)_(m)NR₂₁, orNHC(═O)(CH₂)_(n)NR₂₁; and R₃ is hydrogen, in which R is hydrogen orC₁-C₃ alkyl; R₂₁ is a group selected from the group consisting of:

and m and n are independently an integer of 1 to
 6. 4. The novelcephalosporin derivative of claim 1, which is represented by one of thefollowing chemical formulas, an in vivo hydrolysable ester thereof, anin vivo hydrolysable phosphate ester thereof, an isomer thereof, or apharmaceutically acceptable salt thereof:


5. The novel cephalosporin derivative of claim 1, which is representedby one of the following chemical formulas, an in vivo hydrolysable esterthereof, an in vivo hydrolysable phosphate ester thereof, an isomerthereof, or a pharmaceutically acceptable salt thereof:


6. A pharmaceutical composition comprising the novel cephalosporinderivative according to claim 1, an in vivo hydrolysable ester thereof,an in vivo hydrolysable phosphate ester thereof, an isomer thereof, or apharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier, diluent, adjuvant, or any combination thereof, as aneffective ingredient.
 7. A pharmaceutical composition comprising thenovel cephalosporin derivative according to claim 2, an in vivohydrolysable ester thereof, an in vivo hydrolysable phosphate esterthereof, an isomer thereof, or a pharmaceutically acceptable saltthereof; and a pharmaceutically acceptable carrier, diluent, adjuvant,or any combination thereof, as an effective ingredient.
 8. Apharmaceutical composition comprising the novel cephalosporin derivativeaccording to claim 3, an in vivo hydrolysable ester thereof, an in vivohydrolysable phosphate ester thereof, an isomer thereof, or apharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier, diluent, adjuvant, or any combination thereof, as aneffective ingredient.
 9. A pharmaceutical composition comprising thenovel cephalosporin derivative according to claim 4, an in vivohydrolysable ester thereof, an in vivo hydrolysable phosphate esterthereof, an isomer thereof, or a pharmaceutically acceptable saltthereof; and a pharmaceutically acceptable carrier, diluent, adjuvant,or any combination thereof, as an effective ingredient.
 10. Apharmaceutical composition comprising the novel cephalosporin derivativeaccording to claim 5, an in vivo hydrolysable ester thereof, an in vivohydrolysable phosphate ester thereof, an isomer thereof, or apharmaceutically acceptable salt thereof; and a pharmaceuticallyacceptable carrier, diluent, adjuvant, or any combination thereof, as aneffective ingredient.
 11. An antibiotic treatment method comprisingapplying a pharmaceutically effective amount of the novel cephalosporinderivative of claim 1, an in vivo hydrolysable ester thereof, an in vivohydrolysable phosphate ester thereof, an isomer thereof, or apharmaceutically acceptable salt thereof.
 12. An antibiotic treatmentmethod comprising applying a pharmaceutically effective amount of thenovel cephalosporin derivative of claim 2, an in vivo hydrolysable esterthereof, an in vivo hydrolysable phosphate ester thereof, an isomerthereof, or a pharmaceutically acceptable salt thereof.
 13. Anantibiotic treatment method comprising applying a pharmaceuticallyeffective amount of the novel cephalosporin derivative of claim 3, an invivo hydrolysable ester thereof, an in vivo hydrolysable phosphate esterthereof, an isomer thereof, or a pharmaceutically acceptable saltthereof.
 14. An antibiotic treatment method comprising applying apharmaceutically effective amount of the novel cephalosporin derivativeof claim 4, an in vivo hydrolysable ester thereof, an in vivohydrolysable phosphate ester thereof, an isomer thereof, or apharmaceutically acceptable salt thereof.
 15. An antibiotic treatmentmethod comprising applying a pharmaceutically effective amount of thenovel cephalosporin derivative of claim 5, an in vivo hydrolysable esterthereof, an in vivo hydrolysable phosphate ester thereof, an isomerthereof, or a pharmaceutically acceptable salt thereof.
 16. The methodof claim 1, wherein the bacterial infection results from a Gram-positivebacterium.
 17. The method of claim 1, wherein the bacterial infectionresults from a Gram-negative bacterium.
 18. The method of claim 11,wherein the Gram-positive bacterium is selected from the groupconsisting of Staphylococcus, Enterococcus, Streptococcus and acid-fastbacteria.
 19. The method of claim 11, wherein the Gram-negativebacterium is selected from the group consisting of Pseudomonasaeruginosa, Acinetobacter baumannii, and Klebsiella pneumoniae.